LOCUS       V023217                 9228 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023217
VERSION     V023217
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..3902
                     /label="ITGA5(NM_002205)"
                     /note="ITGA5(NM_002205)"
                     /gene="ITGA5"
     CDS             3915..3980
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4004..4592
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4658..4706
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4908..5336
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5350..5679
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5746..6540
                     /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    6716..6837
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6886..6902)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6910..6926
                     /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(6934..6964)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6979..7000
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7288..7876)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8047..8907)
                     /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(8908..9012)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatccg ccaccatggg gagccggacg ccagagtccc
      781 ctctccacgc cgtgcagctg cgctggggcc cccggcgccg acccccgctg ctgccgctgc
      841 tgttgctgct gctgccgccg ccacccaggg tcgggggctt caacttagac gcggaggccc
      901 cagcagtact ctcggggccc ccgggctcct tcttcggatt ctcagtggag ttttaccggc
      961 cgggaacaga cggggtcagt gtgctggtgg gagcacccaa ggctaatacc agccagccag
     1021 gagtgctgca gggtggtgct gtctacctct gtccttgggg tgccagcccc acacagtgca
     1081 cccccattga atttgacagc aaaggctctc ggctcctgga gtcctcactg tccagctcag
     1141 agggagagga gcctgtggag tacaagtcct tgcagtggtt cggggcaaca gttcgagccc
     1201 atggctcctc catcttggca tgcgctccac tgtacagctg gcgcacagag aaggagccac
     1261 tgagcgaccc cgtgggcacc tgctacctct ccacagataa cttcacccga attctggagt
     1321 atgcaccctg ccgctcagat ttcagctggg cagcaggaca gggttactgc caaggaggct
     1381 tcagtgccga gttcaccaag actggccgtg tggttttagg tggaccagga agctatttct
     1441 ggcaaggcca gatcctgtct gccactcagg agcagattgc agaatcttat taccccgagt
     1501 acctgatcaa cctggttcag gggcagctgc agactcgcca ggccagttcc atctatgatg
     1561 acagctacct aggatactct gtggctgttg gtgaattcag tggtgatgac acagaagact
     1621 ttgttgctgg tgtgcccaaa gggaacctca cttacggcta tgtcaccatc cttaatggct
     1681 cagacattcg atccctctac aacttctcag gggaacagat ggcctcctac tttggctatg
     1741 cagtggccgc cacagacgtc aatggggacg ggctggatga cttgctggtg ggggcacccc
     1801 tgctcatgga tcggacccct gacgggcggc ctcaggaggt gggcagggtc tacgtctacc
     1861 tgcagcaccc agccggcata gagcccacgc ccacccttac cctcactggc catgatgagt
     1921 ttggccgatt tggcagctcc ttgacccccc tgggggacct ggaccaggat ggctacaatg
     1981 atgtggccat cggggctccc tttggtgggg agacccagca gggagtagtg tttgtatttc
     2041 ctgggggccc aggagggctg ggctctaagc cttcccaggt tctgcagccc ctgtgggcag
     2101 ccagccacac cccagacttc tttggctctg cccttcgagg aggccgagac ctggatggca
     2161 atggatatcc tgatctgatt gtggggtcct ttggtgtgga caaggctgtg gtatacaggg
     2221 gccgccccat cgtgtccgct agtgcctccc tcaccatctt ccccgccatg ttcaacccag
     2281 aggagcggag ctgcagctta gaggggaacc ctgtggcctg catcaacctt agcttctgcc
     2341 tcaatgcttc tggaaaacac gttgctgact ccattggttt cacagtggaa cttcagctgg
     2401 actggcagaa gcagaaggga ggggtacggc gggcactgtt cctggcctcc aggcaggcaa
     2461 ccctgaccca gaccctgctc atccagaatg gggctcgaga ggattgcaga gagatgaaga
     2521 tctacctcag gaacgagtca gaatttcgag acaaactctc gccgattcac atcgctctca
     2581 acttctcctt ggacccccaa gccccagtgg acagccacgg cctcaggcca gccctacatt
     2641 atcagagcaa gagccggata gaggacaagg ctcagatctt gctggactgt ggagaagaca
     2701 acatctgtgt gcctgacctg cagctggaag tgtttgggga gcagaaccat gtgtacctgg
     2761 gtgacaagaa tgccctgaac ctcactttcc atgcccagaa tgtgggtgag ggtggcgcct
     2821 atgaggctga gcttcgggtc accgcccctc cagaggctga gtactcagga ctcgtcagac
     2881 acccagggaa cttctccagc ctgagctgtg actactttgc cgtgaaccag agccgcctgc
     2941 tggtgtgtga cctgggcaac cccatgaagg caggagccag tctgtggggt ggccttcggt
     3001 ttacagtccc tcatctccgg gacactaaga aaaccatcca gtttgacttc cagatcctca
     3061 gcaagaatct caacaactcg caaagcgacg tggtttcctt tcggctctcc gtggaggctc
     3121 aggcccaggt caccctgaac ggtgtctcca agcctgaggc agtgctattc ccagtaagcg
     3181 actggcatcc ccgagaccag cctcagaagg aggaggacct gggacctgct gtccaccatg
     3241 tctatgagct catcaaccaa ggccccagct ccattagcca gggtgtgctg gaactcagct
     3301 gtccccaggc tctggaaggt cagcagctcc tatatgtgac cagagttacg ggactcaact
     3361 gcaccaccaa tcaccccatt aacccaaagg gcctggagtt ggatcccgag ggttccctgc
     3421 accaccagca aaaacgggaa gctccaagcc gcagctctgc ttcctcggga cctcagatcc
     3481 tgaaatgccc ggaggctgag tgtttcaggc tgcgctgtga gctcgggccc ctgcaccaac
     3541 aagagagcca aagtctgcag ttgcatttcc gagtctgggc caagactttc ttgcagcggg
     3601 agcaccagcc atttagcctg cagtgtgagg ctgtgtacaa agccctgaag atgccctacc
     3661 gaatcctgcc tcggcagctg ccccaaaaag agcgtcaggt ggccacagct gtgcaatgga
     3721 ccaaggcaga aggcagctat ggcgtcccac tgtggatcat catcctagcc atcctgtttg
     3781 gcctcctgct cctaggtcta ctcatctaca tcctctacaa gcttggattc ttcaaacgct
     3841 ccctcccata tggcaccgcc atggaaaaag ctcagctcaa gcctccagcc acctctgatg
     3901 ccctcgagtc tagagactac aaagaccatg acggtgatta taaagatcat gacatcgact
     3961 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     4021 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4081 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4141 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4201 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4261 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4321 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4381 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4441 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4501 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4561 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     4621 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4681 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4741 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4801 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4861 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     4921 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4981 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5041 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     5101 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5161 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5221 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     5281 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     5341 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5401 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5461 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5521 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5581 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5641 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5701 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     5761 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     5821 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     5881 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     5941 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6001 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6061 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6121 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6181 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6241 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     6301 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6361 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6421 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6481 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6541 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     6601 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6661 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     6721 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6781 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6841 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6901 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6961 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7021 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7081 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7141 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7201 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7261 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7321 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7381 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7441 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7501 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7561 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7621 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7681 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7741 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7801 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7861 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8041 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8101 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8161 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8221 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8281 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8341 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8401 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8461 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8521 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8581 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8641 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8701 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8761 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8821 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8881 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8941 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9001 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     9061 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     9121 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     9181 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
