LOCUS       V034453                 7541 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034453
VERSION     V034453
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3034
                     /label="NCSTN(NM_015331)"
                     /note="NCSTN(NM_015331)"
                     /gene="NCSTN"
     CDS             3044..3061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3108..3332
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3378..3806
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3820..4149
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4216..5010
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5184..5305
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5354..5370)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5378..5394
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5402..5432)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5447..5468
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5756..6341)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6512..7372)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7373..7477)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gctacggcag ggggtggctc tggggctgac ccgggaagtc ggggtctcct
      961 tcgccttctg tctttctgcg tcctactagc aggtttgtgc aggggaaact cagtggagag
     1021 gaagatatat atccccttaa ataaaacagc tccctgtgtt cgcctgctca acgccactca
     1081 tcagattggc tgccagtctt caattagtgg agacacaggg gttatccacg tagtagagaa
     1141 agaggaggac ctacagtggg tattgactga tggccccaac cccccttaca tggttctgct
     1201 ggagagcaag cattttacca gggatttaat ggagaagctg aaagggagaa ccagccgaat
     1261 tgctggtctt gcagtgtcct tgaccaagcc cagtcctgcc tcaggcttct ctcctagtgt
     1321 acagtgccca aatgatgggt ttggtgttta ctccaattcc tatgggccag agtttgctca
     1381 ctgcagagaa atacagtgga attcgctggg caatggtttg gcttatgaag actttagttt
     1441 ccccatcttt cttcttgaag atgaaaatga aaccaaagtc atcaagcagt gctatcaaga
     1501 tcacaacctg agtcagaatg gctcagcacc aaccttccca ctatgtgcca tgcagctctt
     1561 ttcacacatg catgctgtca tcagcactgc cacctgcatg cggcgcagct ccatccaaag
     1621 caccttcagc atcaacccag aaatcgtctg tgaccccctg tctgattaca atgtgtggag
     1681 catgctaaag cctataaata caactgggac attaaagcct gacgacaggg ttgtggttgc
     1741 tgccacccgg ctggatagtc gttccttttt ctggaatgtg gccccagggg ctgaaagcgc
     1801 agtggcttcc tttgtcaccc agctggctgc tgctgaagct ttgcaaaagg cacctgatgt
     1861 gaccaccctg ccccgcaatg tcatgtttgt cttctttcaa ggggaaactt ttgactacat
     1921 tggcagctcg aggatggtct acgatatgga gaagggcaag tttcccgtgc agttagagaa
     1981 tgttgactca tttgtggagc tgggacaggt ggccttaaga acttcattag agctttggat
     2041 gcacacagat cctgtttctc agaaaaatga gtctgtacgg aaccaggtgg aggatctcct
     2101 ggccacattg gagaagagtg gtgctggtgt ccctgctgtc atcctcagga ggccaaatca
     2161 gtcccagcct ctcccaccat cttccctgca gcgatttctt cgagctcgaa acatctctgg
     2221 cgttgttctg gctgaccact ctggtgcctt ccataacaaa tattaccaga gtatttacga
     2281 cactgctgag aacattaatg tgagctatcc cgaatggctg agccctgaag aggacctgaa
     2341 ctttgtaaca gacactgcca aggccctggc agatgtggcc acggtgctgg gacgtgctct
     2401 gtatgagctt gcaggaggaa ccaacttcag cgacacagtt caggctgatc cccaaacggt
     2461 tacccgcctg ctctatgggt tcctgattaa agccaacaac tcatggttcc agtctatcct
     2521 caggcaggac ctaaggtcct acttgggtga cgggcctctt caacattaca tcgctgtctc
     2581 cagccccacc aacaccactt atgttgtaca gtatgccttg gcaaatttga ctggcacagt
     2641 ggtcaacctc acccgagagc agtgccagga tccaagtaaa gtcccaagtg aaaacaagga
     2701 tctgtatgag tactcatggg tccagggccc tttgcattct aatgagacgg accgactccc
     2761 ccggtgtgtg cgttctactg cacgattagc cagggccttg tctcctgcct ttgaactgag
     2821 tcagtggagc tctactgaat actctacatg gactgagagc cgctggaaag atatccgtgc
     2881 ccggatattt ctcatcgcca gcaaagagct tgagttgatc accctgacag tgggcttcgg
     2941 catcctcatc ttctccctca tcgtcaccta ctgcatcaat gccaaagctg atgtcctttt
     3001 cattgctccc cgggagccag gagctgtgtc atacgcggcc gcacatcacc atcatcacca
     3061 ttgactcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3121 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3181 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3241 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     3301 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     3361 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3781 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3841 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3901 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3961 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4021 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4081 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4141 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4201 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4261 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4321 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4381 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4441 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4501 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4561 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4621 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4681 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4741 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4801 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4861 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4921 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4981 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5041 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5101 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5161 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5221 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5281 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     5341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5401 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     6301 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6361 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6421 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6481 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6541 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6601 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6661 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6721 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6781 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6841 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6901 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6961 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7021 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7081 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7141 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7201 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7261 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7321 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7381 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7441 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7501 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
