LOCUS       V025153                 7847 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025153
VERSION     V025153
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..3337
                     /label="ITGB5(NM_002213)"
                     /note="ITGB5(NM_002213)"
                     /gene="ITGB5"
     CDS             3344..3409
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3447..3671
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3717..4145
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4159..4488
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4555..5349
                     /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    5523..5644
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5693..5709)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5717..5733
                     /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(5741..5771)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5786..5807
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6095..6680)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6851..7711)
                     /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(7712..7816)
                     /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 atgccgcggg ccccggcgcc
      961 gctgtacgcc tgcctcctgg ggctctgcgc gctcctgccc cggctcgcag gtctcaacat
     1021 atgcactagt ggaagtgcca cctcatgtga agaatgtctg ctaatccacc caaaatgtgc
     1081 ctggtgctcc aaagaggact tcggaagccc acggtccatc acctctcggt gtgatctgag
     1141 ggcaaacctt gtcaaaaatg gctgtggagg tgagatagag agcccagcca gcagcttcca
     1201 tgtcctgagg agcctgcccc tcagcagcaa gggttcgggc tctgcaggct gggacgtcat
     1261 tcagatgaca ccacaggaga ttgccgtgaa cctccggccc ggtgacaaga ccaccttcca
     1321 gctacaggtt cgccaggtgg aggactatcc tgtggacctg tactacctga tggacctctc
     1381 cctgtccatg aaggatgact tggacaatat ccggagcctg ggcaccaaac tcgcggagga
     1441 gatgaggaag ctcaccagca acttccggtt gggatttggg tcttttgttg ataaggacat
     1501 ctctcctttc tcctacacgg caccgaggta ccagaccaat ccgtgcattg gttacaagtt
     1561 gtttccaaat tgcgtcccct cctttgggtt ccgccatctg ctgcctctca cagacagagt
     1621 ggacagcttc aatgaggaag ttcggaaaca gagggtgtcc cggaaccgag atgcccctga
     1681 ggggggcttt gatgcagtac tccaggcagc cgtctgcaag gagaagattg gctggcgaaa
     1741 ggatgcactg catttgctgg tgttcacaac agatgatgtg ccccacatcg cattggatgg
     1801 aaaattggga ggcctggtgc agccacacga tggccagtgc cacctgaacg aggccaacga
     1861 gtacactgca tccaaccaga tggactatcc atcccttgcc ttgcttggag agaaattggc
     1921 agagaacaac atcaacctca tctttgcagt gacaaaaaac cattatatgc tgtacaagaa
     1981 ttttacagcc ctgatacctg gaacaacggt ggagatttta gatggagact ccaaaaatat
     2041 tattcaactg attattaatg catacaatag tatccggtct aaagtggagt tgtcagtctg
     2101 ggatcagcct gaggatctta atctcttctt tactgctacc tgccaagatg gggtatccta
     2161 tcctggtcag aggaagtgtg agggtctgaa gattggggac acggcatctt ttgaagtatc
     2221 attggaggcc cgaagctgtc ccagcagaca cacggagcat gtgtttgccc tgcggccggt
     2281 gggattccgg gacagcctgg aggtgggggt cacctacaac tgcacgtgcg gctgcagcgt
     2341 ggggctggaa cccaacagcg ccaggtgcaa cgggagcggg acctatgtct gcggcctgtg
     2401 tgagtgcagc cccggctacc tgggcaccag gtgcgagtgc caggatgggg agaaccagag
     2461 cgtgtaccag aacctgtgcc gggaggcaga gggcaagcca ctgtgcagcg ggcgtgggga
     2521 ctgcagctgc aaccagtgct cctgcttcga gagcgagttt ggcaagatct atgggccttt
     2581 ctgtgagtgc gacaacttct cctgtgccag gaacaaggga gtcctctgct caggccatgg
     2641 cgagtgtcac tgcggggaat gcaagtgcca tgcaggttac atcggggaca actgtaactg
     2701 ctcgacagac atcagcacat gccggggcag agatggccag atctgcagcg agcgtgggca
     2761 ctgtctctgt gggcagtgcc aatgcacgga gccgggggcc tttggggaga tgtgtgagaa
     2821 gtgccccacc tgcccggatg catgcagcac caagagagat tgcgtcgagt gcctgctgct
     2881 ccactctggg aaacctgaca accagacctg ccacagccta tgcagggatg aggtgatcac
     2941 atgggtggac accatcgtga aagatgacca ggaggctgtg ctatgtttct acaaaaccgc
     3001 caaggactgc gtcatgatgt tcacctatgt ggagctcccc agtgggaagt ccaacctgac
     3061 cgtcctcagg gagccagagt gtggaaacac ccccaacgcc atgaccatcc tcctggctgt
     3121 ggtcggtagc atcctccttg ttgggcttgc actcctggct atctggaagc tgcttgtcac
     3181 catccacgac cggagggagt ttgcaaagtt tcagagcgag cgatccaggg cccgctatga
     3241 aatggcttca aatccattat acagaaagcc tatctccacg cacactgtgg acttcacctt
     3301 caacaagttc aacaaatcct acaatggcac tgtggacctc gaggactaca aagaccatga
     3361 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     3421 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3481 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3541 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3601 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3661 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3721 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3781 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3841 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3901 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3961 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4021 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4081 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4141 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4201 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4261 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4321 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4381 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4441 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4501 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4561 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4621 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4681 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4741 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4801 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4861 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4921 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4981 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5041 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     5101 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5161 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5221 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5281 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5341 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5401 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5461 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5521 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5581 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5641 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5701 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5761 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5821 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5881 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5941 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6001 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6601 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6661 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6721 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6781 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6841 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6901 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6961 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7021 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7081 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7141 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7201 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7261 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7321 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7381 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7441 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7501 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7561 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7621 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7681 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7741 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
