LOCUS       Exported                7702 bp ds-DNA     circular SYN 03-OCT-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    niCAR in pET32a-LIC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  AUTHORS   Will Finnigan
  TITLE     Direct Submission
  JOURNAL   Exported 3 Oct 2016 from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7702
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..910
                     /note="nCAR 8"
     misc_feature    911..1674
                     /note="nCAR out Fwd"
     promoter        complement(1201..1278)
                     /gene="lacI"
                     /note="lacI promoter"
     promoter        1587..1605
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1606..1630
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     misc_feature    1635..2388
                     /note="t7 Fwd"
     CDS             1674..1676
                     /codon_start=1
                     /note="Start"
                     /translation="M"
     CDS             1680..1697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             1704..5228
                     /codon_start=1
                     /note="nCAR"
                     /translation="MAVDSPDERLQRRIAQLFAEDEQVKAARPLEAVSAAVSAPGMRLA
                     QIAATVMAGYADRPAAGQRAFELNTDDATGRTSLRLLPRFETITYRELWQRVGEVAAAW
                     HHDPENPLRAGDFVALLGFTSIDYATLDLADIHLGAVTVPLQASAAVSQLIAILTETSP
                     RLLASTPEHLDAAVECLLAGTTPERLVVFDYHPEDDDQRAAFESARRRLADAGSLVIVE
                     TLDAVRARGRDLPAAPLFVPDTDDDPLALLIYTSGSTGTPKGAMYTNRLAATMWQGNSM
                     LQGNSQRVGINLNYMPMSHIAGRISLFGVLARGGTAYFAAKSDMSTLFEDIGLVRPTEI
                     FFVPRVCDMVFQRYQSELDRRSVAGADLDTLDREVKADLRQNYLGGRFLVAVVGSAPLA
                     AEMKTFMESVLDLPLHDGYGSTEAGASVLLDNQIQRPPVLDYKLVDVPELGYFRTDRPH
                     PRGELLLKAETTIPGYYKRPEVTAEIFDEDGFYKTGDIVAELEHDRLVYVDRRNNVLKL
                     SQGEFVTVAHLEAVFASSPLIRQIFIYGSSERSYLLAVIVPTDDALRGRDTATLKSALA
                     ESIQRIAKDANLQPYEIPRDFLIETEPFTIANGLLSGIAKLLRPNLKERYGAQLEQMYT
                     DLATGQADELLALRREAADLPVLETVSRAAKAMLGVASADMRPDAHFTDLGGDSLSALS
                     FSNLLHEIFGVEVPVGVVVSPANELRDLANYIEAERNSGAKRPTFTSVHGGGSEIRAAD
                     LTLDKFIDARTLAAADSIPHAPVPAQTVLLTGANGYLGRFLCLEWLERLDKTGGTLICV
                     VRGSDAAAARKRLDSAFDSGDPGLLEHYQQLAARTLEVLAGDIGDPNLGLDDATWQRLA
                     ETVDLIVHPAALVNHVLPYTQLFGPNVVGTAEIVRLAITARRKPVTYLSTVGVADQVDP
                     AEYQEDSDVREMSAVRVVRESYANGYGNSKWAGEVLLREAHDLCGLPVAVFRSDMILAH
                     SRYAGQLNVQDVFTRLILSLVATGIAPYSFYRTDADGNRQRAHYDGLPADFTAAAITAL
                     GIQATEGFRTYDVLNPYDDGISLDEFVDWLVESGHPIQRITDYSDWFHRFETAIRALPE
                     KQRQASVLPLLDAYRNPCPAVRGAILPAKEFQAAVQTAKIGPEQDIPHLSAPLIDKYVS
                     DLELLQLL"
     misc_feature    2311..3176
                     /note="nCAR 7"
     misc_feature    2436..2826
                     /note="nCAR 1"
     misc_feature    2827..3657
                     /note="nCAR 6"
     misc_feature    3795..4459
                     /note="nCAR out"
     misc_feature    4395..5218
                     /note="nCAR 5"
     misc_feature    4460..5288
                     /note="T7 Rev"
     CDS             5241..5258
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     misc_feature    5259..6100
                     /note="nCAR 3"
     misc_feature    5323..6100
                     /note="nCAR 2"
     terminator      5325..5372
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5409..5864
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5890..5994
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             5995..6855
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    6047..6950
                     /note="nCAR 4"
     rep_origin      7026..7614
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
       61 aacgcaacga gtcagtgacc gaggaagcgg aaaaaccgcc tgaagcggta ttttctcctt
      121 accccccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      181 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt tttcttttca ccagtgagac
      241 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
      301 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
      361 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
      421 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
      481 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
      541 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
      601 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
      661 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
      721 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
      781 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
      841 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
      901 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
      961 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     1021 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     1081 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     1141 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     1201 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     1261 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     1321 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     1381 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     1441 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     1501 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     1561 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     1621 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgagcc
     1681 atcatcatca tcatcatggc accatggcgg tggatagccc ggatgaacgt ctgcagcgtc
     1741 gtattgcgca gctgtttgcg gaagatgaac aggtgaaagc agcacgcccg ctggaagcgg
     1801 ttagcgcagc ggtgagcgca ccgggtatgc gtctggccca gattgcggcg accgtgatgg
     1861 cgggctatgc ggatcgtccg gcagcgggtc agcgtgcgtt tgaactgaac accgatgatg
     1921 cgaccggccg taccagcctg cgtctgctgc cgcgttttga aaccattacc tatcgtgaac
     1981 tgtggcagcg tgtgggtgaa gttgcggcag cgtggcatca cgatccggaa aatccgctgc
     2041 gtgcgggcga ttttgtggcg ctgctgggct ttaccagcat tgattatgcg accctggatc
     2101 tggccgatat tcatctgggc gcggtgaccg ttccgctgca ggcgagcgca gcagtcagcc
     2161 aactgattgc gattctgacc gaaacgagtc cgcgcctgct ggcatctacc ccggaacatc
     2221 tggatgcggc ggtggaatgt ctgctggcag gtacgacgcc ggaacgcctg gtggtgtttg
     2281 attatcatcc ggaagatgat gatcagcgtg cggcgtttga aagcgcgcgt cgtcgtctgg
     2341 ccgatgcggg cagcctggtg attgtggaaa ccctggatgc ggtgcgtgcg cgtggtcgtg
     2401 atctgccggc tgctccgctg tttgtgccgg ataccgatga tgatccgctg gccctgctga
     2461 tttatacctc tggtagcacg ggtaccccga aaggcgccat gtataccaac cgcctggcag
     2521 caacgatgtg gcaaggtaac agcatgctgc agggcaatag ccagcgtgtg ggcattaacc
     2581 tgaactatat gccgatgagc catattgcgg gccgtattag cctgtttggc gtgctggccc
     2641 gtggtggcac cgcgtatttt gcggcgaaaa gcgatatgag caccctgttt gaagatattg
     2701 gcctggtgcg tccgaccgaa attttttttg tgccgcgtgt gtgcgatatg gtgtttcagc
     2761 gttatcagag cgaactggat cgtcgtagcg tggcgggtgc ggatctggat accctggatc
     2821 gtgaagtgaa agcggatctg cgtcagaact atctgggcgg tcgttttctg gtggcggtgg
     2881 tgggtagcgc accgctggcc gcggaaatga aaacctttat ggaaagcgtg ctggatctgc
     2941 cgctgcatga tggctatggc agcaccgaag cgggtgcgag cgtgctgctg gataaccaga
     3001 ttcagcgtcc gccggtgctg gattataaac tggtggacgt cccggaactg ggctattttc
     3061 gtaccgatcg tccgcatccg cgtggcgaac tgctgctgaa agcggaaacc accattccgg
     3121 gctattataa acgtccggaa gtgaccgcgg aaatttttga tgaagatggc ttctataaaa
     3181 ccggcgatat tgtggcggaa ctggaacatg atcgtctggt gtatgtggat cgtcgcaaca
     3241 acgtgctgaa actgagccag ggcgaatttg tgaccgtggc gcatctggaa gcggtgtttg
     3301 cgagcagccc gctgattcgt cagattttta tctacggctc tagtgaacgc tcttatctgc
     3361 tggcagtgat tgtgccgacc gatgatgccc tgcgtggccg tgataccgcg accctgaaaa
     3421 gcgcgctggc cgaaagcatt cagcgtattg cgaaagatgc gaacctgcag ccgtatgaaa
     3481 ttccgcgtga ttttctgatt gaaaccgaac cgttcaccat tgcgaacggc ctgctgtctg
     3541 gcattgcgaa actgctgcgt ccgaacctga aagaacgtta tggcgcgcag ctggaacaaa
     3601 tgtataccga tctggccacc ggccaggcgg atgaactgct ggccctgcgt cgtgaagcgg
     3661 cggatctgcc ggttctggaa accgttagcc gtgcggcgaa agccatgctg ggtgtggcga
     3721 gcgcggatat gcgtccggat gcgcatttta ccgatctggg cggcgatagc ctgagcgccc
     3781 tgagctttag caacctgctg catgaaattt ttggcgtgga agtgccggtg ggtgtggttg
     3841 tgagcccggc aaacgaactg cgtgacctgg ccaactatat tgaagcggaa cgtaacagcg
     3901 gcgcgaaacg tccgaccttt accagcgtgc atggcggcgg tagcgaaatt cgtgcggccg
     3961 atctgaccct ggataaattt attgatgcgc gtaccctggc cgcagcggat agcattccgc
     4021 atgcaccggt tccggcacag accgtcctgc tgacgggcgc aaatggctat ctgggccgtt
     4081 ttctgtgcct ggaatggctg gaacgtctgg ataaaaccgg tggcaccctg atttgcgtgg
     4141 tgcgtggcag cgatgcggcg gcagcccgta aacgcctgga tagcgcgttt gatagcggcg
     4201 atccgggcct gctggaacat tatcagcagc tggccgcacg caccctggaa gttctggccg
     4261 gtgatattgg cgatccgaac ctgggcctgg atgatgccac ctggcagcgt ctggccgaaa
     4321 ccgtggatct gattgtgcac ccggctgctc tggtgaatca tgtgctgccg tatacccagc
     4381 tgtttggccc gaacgttgtg ggcaccgcgg aaatcgttcg tctggctatt accgcgcgtc
     4441 gtaaaccggt gacctatctg agcaccgtgg gcgtggcgga tcaggttgat ccggcggaat
     4501 atcaggaaga tagcgacgtc cgcgaaatga gcgcagtccg cgtcgttcgc gaaagttatg
     4561 caaacggtta tggtaacagc aaatgggcgg gtgaagtgct gctgcgtgaa gcgcatgatc
     4621 tgtgcggtct gccggtggcg gtgtttcgta gcgatatgat tctggcccat agccgttatg
     4681 cgggccagct gaacgtgcag gatgtgttta cccgtctgat tctgagcctg gtggcgaccg
     4741 gcattgcgcc gtatagcttt tatcgcaccg atgcggatgg caaccgtcag cgtgcgcatt
     4801 atgatggcct gccggcggat tttaccgcag cagctatcac cgcactgggc attcaggcga
     4861 ccgaaggctt tcgtacctat gatgtgctga atccgtatga tgatggcatt agcctggatg
     4921 aatttgtcga ttggctggtc gaatctggtc acccgattca gcgcattacc gattatagcg
     4981 attggtttca ccgctttgaa accgcgattc gtgcgctgcc ggaaaaacag cgtcaggcga
     5041 gcgttctgcc gctgctggat gcgtatcgta atccgtgtcc ggcggtccgt ggtgcaattc
     5101 tgccggcgaa agaatttcag gcggcggtgc agaccgcgaa aattggcccg gaacaggata
     5161 ttccgcatct gagcgcaccg ctgattgata aatatgtgag cgatctggaa ctgctgcagc
     5221 tgctgtaaga attcctcgag caccaccacc accaccactg agatccggct gctaacaaag
     5281 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5341 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     5401 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5761 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5821 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5881 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5941 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6001 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6061 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6121 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6181 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6241 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6301 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6361 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6421 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6481 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6541 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6601 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6661 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6721 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6781 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6841 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6901 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6961 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7021 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7081 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7141 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     7201 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7261 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7321 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7381 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7441 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7501 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7561 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7621 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7681 cctgcgttat cccctgattc tg
//
