LOCUS       V019815                 6224 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019815
VERSION     V019815
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..1714
                     /label="SNAP29(NM_004782)"
                     /note="SNAP29(NM_004782)"
                     /gene="SNAP29"
     CDS             1721..1786
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    1824..2048
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2094..2522
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2536..2865
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2932..3726
                     /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    3900..4021
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4070..4086)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4094..4110
                     /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(4118..4148)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4163..4184
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4472..5057)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5228..6088)
                     /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(6089..6193)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgtcagctt accctaaaag
      961 ctacaatccg ttcgacgacg acggggagga cgaaggcgcc cggccggccc cttggaggga
     1021 cgcccgagac ctccccgacg ggcccgacgc gcccgcggac aggcagcagt acttgcggca
     1081 ggaggtcctc cgcagggctg aggccacggc cgccagcacc agcaggtccc tggccctcat
     1141 gtacgagtcc gagaaggttg gggtcgcctc ttccgaggag ctcgcccgtc agcgaggagt
     1201 cctggagcgc acagagaaga tggtggacaa gatggaccaa gatttgaaga tcagccagaa
     1261 acacatcaat agcattaaga gcgtgtttgg ggggctggtc aattacttca aatccaaacc
     1321 agtagagacc ccacctgaac agaatggcac cctcacctcc cagcccaaca acagattgaa
     1381 agaagctata agtacaagta aagaacagga agcaaagtac caggccagcc acccaaacct
     1441 tagaaagctg gatgatacag accctgtccc cagaggggct ggttctgcca tgagtactga
     1501 tgcttaccca aagaacccac accttcgagc ctatcaccag aagatcgaca gcaacctaga
     1561 tgagctgtcc atgggactgg gtcgtctgaa ggacatagcc ctggggatgc agacagaaat
     1621 tgaggagcaa gatgacattc ttgaccggct gacaaccaaa gtggacaagt tagatgtcaa
     1681 cataaaaagc acagaaagaa aagttcgaca actcctcgag gactacaaag accatgacgg
     1741 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt
     1801 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     1861 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     1921 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     1981 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2041 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2101 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2161 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2221 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2281 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2341 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2401 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2461 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2521 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2581 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2641 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2701 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2761 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2821 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     2881 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2941 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3001 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3061 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3121 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3181 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3241 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3301 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3361 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3421 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3481 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3541 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3601 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3661 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3721 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3781 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3841 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3901 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3961 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4021 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4081 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4141 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4201 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4261 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4321 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4381 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4441 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4501 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4561 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4621 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4681 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4741 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4801 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4861 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4921 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4981 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5041 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5101 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5161 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5221 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5281 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5341 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5401 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5461 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5521 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5581 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5641 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5701 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5761 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5821 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5881 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5941 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6001 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6061 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6121 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6181 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
