LOCUS       V020149                 8207 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020149
VERSION     V020149
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..2887
                     /label="EXOC7(NM_001145297)"
                     /note="EXOC7(NM_001145297)"
                     /gene="EXOC7"
     CDS             2894..2959
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2983..3571
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3637..3685
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3887..4315
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4329..4658
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4725..5519
                     /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    5695..5816
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5865..5881)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5889..5905
                     /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(5913..5943)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5958..5979
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6267..6855)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7026..7886)
                     /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(7887..7991)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgattcc cccacaggag gcatccgctc gacggcggga
      721 gattgaggac aagctgaagc aggaggagga gactctgtcc ttcatccgag acagcctgga
      781 gaagagcgac cagctcacta agaacatggt gtctatctta tcatcctttg agagccgcct
      841 tatgaagctg gagaactcca tcatccctgt gcacaagcag acggagaatc tgcagcggct
      901 gcaggagaat gttgagaaga cgctgtcctg cctggaccat gtcatcagct actaccatgt
      961 ggccagtgac actgagaaga tcatcagaga gggccccaca ggtaggctgg aagagtacct
     1021 gggaagcatg gccaagattc agaaggcagt ggagtatttc caggacaaca gcccagacag
     1081 cccggaactc aacaaagtga aactgctctt tgagcgcggg aaggaggccc tggagtccga
     1141 atttcgcagc ctgatgacgc ggcacagtaa ggtcgtctcg cccgtgctca tcttggatct
     1201 gatcagtggt gacgatgatc tggaggccca ggaggacgtg accctggagc acctgcccga
     1261 gagcgtgctc caggatgtca ttcgcatctc ccgctggctg gtggaatatg gccgcaacca
     1321 agatttcatg aacgtctact accagatacg ctccagccag ctggaccgct ccatcaaagg
     1381 actgaaggag catttccata agagcagttc ttcctctggg gttccctact cccctgctat
     1441 ccccaacaag aggaaagaca cacctaccaa gaagccagtc aagcggccag ggacgatccg
     1501 taaggctcag aaccttctga aacagtattc ccagcatggt ctagatggga aaaagggggg
     1561 ctctaacctc attcctctgg aagggcttct tccctgtacc ccaaggggcg gcctgcctgg
     1621 cccctggata aacgctgctt gtgtttgtgc ggccgacatc tcgccaggtc acgagcatga
     1681 tttccgagtt aagcacctgt ccgaggcctt gaacgacaag cacgggccgc tggccgggag
     1741 agatgacatg ctggacgtgg agaccgatgc ctacatccac tgcgtcagtg ccttcgtcaa
     1801 gctggcgcag agcgagtacc agctgctggc cgacatcatc cccgagcacc accagaagaa
     1861 gaccttcgac tccctgatac aggatgccct ggatgggctg atgcttgaag gggagaacat
     1921 cgtgtctgct gcccggaagg ccattgtgcg acacgacttc tccacggtgc tcaccgtctt
     1981 ccccatcctg cgacacctca agcagaccaa gcctgagttt gaccaggtgc tccagggcac
     2041 ggctgccagc acaaagaaca agctgcctgg cctcatcaca tccatggaga ccatcggtgc
     2101 caaagcgctg gaggacttcg cagacaacat caagaatgac ccggacaagg agtacaacat
     2161 gccgaaggac ggcaccgtac acgagctcac cagcaatgcc atcctcttcc tgcagcagct
     2221 tttggacttc caggagacgg caggcgccat gctggcctcc caagagacca gctcttcggc
     2281 caccagctac agctctgagt tcagcaagcg gctgctaagc acctatatct gtaaagtgct
     2341 gggcaacctg cagttgaact tgctgagcaa gtccaaggtg tacgaggacc cagctctgag
     2401 cgccatcttc ctgcacaaca actacaatta catcctcaag tccctggaga agtctgaact
     2461 gatccagctg gtggcagtga cacagaagac tgctgagcgc tcctaccggg agcacattga
     2521 gcagcagatc cagacctacc agcgcagctg gttaaaggtg actgattaca tcgcagagaa
     2581 gaatctacct gtgttccagc cgggagtcaa gctccgggac aaggagcggc agattatcaa
     2641 ggagcgtttt aagggcttca atgatggcct cgaagaactg tgcaaaatcc agaaggcctg
     2701 ggctattcca gacacagagc agagggacag gattcgccag gcccagaaga ccattgtcaa
     2761 ggagacctac ggggcctttc tacagaagtt tggcagcgtg cccttcacca agaacccgga
     2821 gaagtacatc aagtacgggg tggagcaggt gggcgacatg atcgatcgcc ttttcgacac
     2881 ctctgccctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     2941 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     3001 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3061 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3121 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3181 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3241 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3301 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3361 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     3421 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3481 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3541 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     3601 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3661 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     3721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     3781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     3841 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     3901 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3961 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4021 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     4081 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4141 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4201 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4261 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4321 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4441 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4501 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4561 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4621 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4681 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4741 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4801 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4861 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4921 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4981 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5041 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5101 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5161 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5221 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5281 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5341 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5401 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     5461 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     5521 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     5581 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     5641 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     5701 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5761 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5821 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6001 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6061 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6121 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6181 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6241 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6301 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6361 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     6421 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     6481 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6541 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6601 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6661 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     6721 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6781 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6841 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6901 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6961 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7021 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7081 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7141 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7201 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7261 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7321 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7381 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7441 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7501 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7561 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7621 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7681 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7741 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7801 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7861 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7921 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7981 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8041 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8101 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8161 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
