LOCUS       V039744                 7507 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039744
VERSION     V039744
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             964..2997
                     /label="GAS6(NM_000820)"
                     /note="GAS6(NM_000820)"
                     /gene="GAS6"
     CDS             3004..3069
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3107..3331
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3377..3805
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3819..4148
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4215..5009
                     /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    5183..5304
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5353..5369)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5377..5393
                     /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(5401..5431)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5446..5467
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5755..6340)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6511..7371)
                     /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(7372..7476)
                     /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 atccactagt ccagtgtggt ggaattcgcc
      961 accatggccc cttcgctctc gcccgggccc gccgccctgc gccgcgcgcc gcagctgctg
     1021 ctgctgctgc tggccgcgga gtgcgcgctt gccgcgctgt tgccggcgcg cgaggccacg
     1081 cagttcctgc ggcccaggca gcgccgcgcc tttcaggtct tcgaggaggc caagcagggc
     1141 cacctggaga gggagtgcgt ggaggagctg tgcagccgcg aggaggcgcg ggaggtgttc
     1201 gagaacgacc ccgagacgga ttatttttac ccaagatact tagactgcat caacaagtat
     1261 gggtctccgt acaccaaaaa ctcaggcttc gccacctgcg tgcaaaacct gcctgaccag
     1321 tgcacgccca acccctgcga taggaagggg acccaagcct gccaggacct catgggcaac
     1381 ttcttctgcc tgtgtaaagc tggctggggg ggccggctct gcgacaaaga tgtcaacgaa
     1441 tgcagccagg agaacggggg ctgcctccag atctgccaca acaagccggg tagcttccac
     1501 tgttcctgcc acagcggctt cgagctctcc tctgatggca ggacctgcca agacatagac
     1561 gagtgcgcag actcggaggc ctgcggggag gcgcgctgca agaacctgcc cggctcctac
     1621 tcctgcctct gtgacgaggg ctttgcgtac agctcccagg agaaggcttg ccgagatgtg
     1681 gacgagtgtc tgcagggccg ctgtgagcag gtctgcgtga actccccagg gagctacacc
     1741 tgccactgtg acgggcgtgg gggcctcaag ctgtcccagg acatggacac ctgtgaggac
     1801 atcttgccgt gcgtgccctt cagcgtggcc aagagtgtga agtccttgta cctgggccgg
     1861 atgttcagtg ggacccccgt gatccgactg cgcttcaaga ggctgcagcc caccaggctg
     1921 gtagctgagt ttgacttccg gacctttgac cccgagggca tcctcctctt tgccggaggc
     1981 caccaggaca gcacctggat cgtgctggcc ctgagagccg gccggctgga gctgcagctg
     2041 cgctacaacg gtgtcggccg tgtcaccagc agcggcccgg tcatcaacca tggcatgtgg
     2101 cagacaatct ctgttgagga gctggcgcgg aatctggtca tcaaggtcaa cagggatgct
     2161 gtcatgaaaa tcgcggtggc cggggacttg ttccaaccgg agcgaggact gtatcatctg
     2221 aacctgaccg tgggaggtat tcccttccat gagaaggacc tcgtgcagcc tataaaccct
     2281 cgtctggatg gctgcatgag gagctggaac tggctgaacg gagaagacac caccatccag
     2341 gaaacggtga aagtgaacac gaggatgcag tgcttctcgg tgacggagag aggctctttc
     2401 taccccggga gcggcttcgc cttctacagc ctggactaca tgcggacccc tctggacgtc
     2461 gggactgaat caacctggga agtagaagtc gtggctcaca tccgcccagc cgcagacaca
     2521 ggcgtgctgt ttgcgctctg ggcccccgac ctccgtgccg tgcctctctc tgtggcactg
     2581 gtagactatc actccacgaa gaaactcaag aagcagctgg tggtcctggc cgtggagcat
     2641 acggccttgg ccctaatgga gatcaaggtc tgcgacggcc aagagcacgt ggtcaccgtc
     2701 tcgctgaggg acggtgaggc caccctggag gtggacggca ccaggggcca gagcgaggtg
     2761 agcgccgcgc agctgcagga gaggctggcc gtgctcgaga ggcacctgcg gagccccgtg
     2821 ctcacctttg ctggcggcct gccagatgtg ccggtgactt cagcgccagt caccgcgttc
     2881 taccgcggct gcatgacact ggaggtcaac cggaggctgc tggacctgga cgaggcggcg
     2941 tacaagcaca gcgacatcac ggcccactcc tgcccccccg tggagcccgc cgcagccctc
     3001 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     3061 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     3121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3721 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4081 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4141 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4201 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4981 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5041 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5101 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5161 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5281 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5341 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5401 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5461 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     6301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7501 tgacgtc
//
