LOCUS       V041767                 9483 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041767
VERSION     V041767
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..4188
                     /label="ITGA8(NM_003638)"
                     /note="ITGA8(NM_003638)"
                     /gene="ITGA8"
     polyA_signal    4242..4864
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        4906..5203
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5257..6051
                     /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    6703..6837
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(6843..6861)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6875..6891)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6899..6915
                     /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(6923..6953)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6968..6989
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7277..7865)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8036..8896)
                     /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(8897..9001)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      9028..9456
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtcgcccgg ggccagccgc
     1021 ggtccccggg gaagccaggc gccgctgatc gcgcccctct gctgcgccgc ggccgcgctg
     1081 gggatgttgc tgtggtcccc cgcctgtcag gcgttcaacc tggacgtgga aaagctcaca
     1141 gtgtacagcg gccccaaggg cagctacttc ggctacgccg tggacttcca catacccgac
     1201 gcccgcacag cgagtgtctt ggtgggggcg cccaaagcca acaccagcca gcccgatatc
     1261 gtggaagggg gagccgtcta ttactgtcct tggcccgcgg aggggtctgc gcagtgcagg
     1321 cagataccgt ttgacaccac caacaacaga aagatcagag ttaatggaac caaagaacct
     1381 atcgagttca aatccaatca gtggtttgga gcaacagtga aagctcacaa aggaaaagtt
     1441 gtggcctgtg ctcctttata tcactggaga actcttaaac cgacaccaga aaaggaccca
     1501 gttggcacct gctatgtagc aattcagaac ttcagcgcct atgccgagtt ctctccttgc
     1561 cggaacagca atgctgatcc ggaaggccag ggttactgcc aagcaggatt tagtctggat
     1621 ttttataaga atggagacct tattgtggga ggacctggga gtttctactg gcaaggacaa
     1681 gtgatcactg ccagtgttgc agatatcatt gcaaattact cattcaagga tatcctcagg
     1741 aaactggcag gagaaaagca gacggaagtg gctccagctt cctatgatga cagttacctt
     1801 ggatactcag ttgctgctgg ggagtttact ggggattctc agcaagaatt ggttgctgga
     1861 attccaagag gagcacagaa ttttggatat gtttccatca ttaactctac ggatatgacg
     1921 tttattcaga atttcacggg agaacagatg gcatcttatt ttggatatac cgttgtcgta
     1981 tcagatgtta acagtgatgg actggatgat gtcctggttg gggcacctct ctttatggaa
     2041 cgtgaatttg agagcaaccc cagagaagta gggcaaatct acctgtattt gcaagtgagc
     2101 tctctcctct tcagagaccc ccagatcctc actggcaccg agacgtttgg gagattcggt
     2161 agtgctatgg cacacttagg agacctgaac caagatggat acaatgacat tgccatcgga
     2221 gtgccttttg caggcaagga tcaaagaggc aaagtgctca tttataatgg gaacaaagat
     2281 ggcttaaaca ccaagccttc ccaagttctg caaggagtgt gggcctcaca tgctgtccct
     2341 tccggatttg gctttacttt aagaggagat tcagacatag acaagaatga ttacccagat
     2401 ttgattgtgg gtgcatttgg aacaggaaaa gtcgctgttt acagagcaag accggttgtg
     2461 actgtagatg cccagcttct gctgcaccca atgattatca atcttgaaaa taaaacttgc
     2521 caggttccag actctatgac atctgctgcc tgcttttctt taagagtatg tgcatctgtc
     2581 acaggccaga gcattgcaaa cacaatagtc ttgatggcag aggtgcaatt agattccctg
     2641 aaacagaaag gagctattaa acggacgctc ttccttgata accatcaggc tcatcgcgtc
     2701 ttccctcttg tgataaaaag gcagaaatcc caccagtgcc aggatttcat cgtttacctt
     2761 cgagatgaaa ctgaattccg agataaatta tctccaatca acattagttt gaattacagt
     2821 ttggacgaat ccacctttaa agaaggcctg gaagtgaaac caatattgaa ctactacaga
     2881 gaaaacattg ttagtgaaca ggctcacatt ctggtggact gtggagaaga caatctgtgt
     2941 gttcctgact tgaagctgtc ggctagacca gataagcatc aggtaatcat tggagatgaa
     3001 aatcacctta tgctcataat aaatgcaaga aatgaagggg aaggagcata tgaagctgaa
     3061 ctctttgtaa tgataccaga agaggcagat tatgttggaa tcgaacgcaa caacaaggga
     3121 tttcgaccac tgagctgtga gtacaagatg gaaaatgtaa ccaggatggt ggtgtgtgac
     3181 cttgggaacc ctatggtgtc tggaacaaat tattccctgg gcctccgatt tgcagttcca
     3241 cgtcttgaga aaacaaacat gagcattaac ttcgatctcc aaatcagaag ttccaacaag
     3301 gacaatccag acagcaattt tgtgagcctg caaatcaaca tcactgctgt agcgcaggtg
     3361 gaaataagag gagtgtcaca ccctccgcag attgttctgc ccattcataa ctgggaacca
     3421 gaagaggagc cccacaaaga ggaggaggtt ggaccattgg tggaacatat ttatgagctg
     3481 cacaatattg gaccaagtac catcagtgac accatcctgg aggtgggctg gcctttctct
     3541 gcccgggatg aatttcttct ctatattttc catattcaaa ctctgggacc tctgcagtgc
     3601 caaccaaatc ctaatatcaa tccacaggat ataaagcctg ctgcctcccc agaggacacc
     3661 cctgagctca gcgccttttt gcgaaactct actattcctc atcttgtcag gaagagggat
     3721 gtacatgtgg tcgaattcca cagacagagc cctgcaaaaa tactgaattg tacaaatatc
     3781 gagtgtttac aaatctcctg tgcagtggga cgactcgaag gaggagaaag cgcagtcctg
     3841 aaagtcaggt cacgattatg ggcccacacc ttcctccaga gaaaaaatga tccctatgct
     3901 cttgcatccc tggtgtcctt tgaagttaag aagatgcctt atacagatca gccagcaaaa
     3961 ctcccagaag gaagcatagt aattaagaca tcagttattt gggcaactcc gaatgtttcc
     4021 ttctcaatcc cattatgggt aataatacta gcaatacttc ttggattgtt ggttctcgcc
     4081 attttaacct tagctttatg gaagtgtgga ttctttgaca gagccagacc tcctcaggag
     4141 gacatgaccg acagggaaca gctgacaaat gacaagaccc ctgaggcatg agaattcatc
     4201 gatagatctg atatcggtac cagtcgactc tagaggatcc cgggtggcat ccctgtgacc
     4261 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     4321 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     4381 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     4441 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     4501 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     4561 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     4621 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4681 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc
     4741 agcaggagga cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt
     4801 tgagttgctt gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg
     4861 aattgggtac gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4921 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4981 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5041 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     5101 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     5161 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggaatt gatcagcttg
     5221 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5281 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5341 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5401 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5461 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5521 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5581 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5641 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5701 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5761 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5821 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5881 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5941 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6001 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6061 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6121 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6181 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     6241 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     6301 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     6361 aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga
     6421 tacagaacga attgcttgca ggcatctcat gagtgtgtct tcccgttttc cgcctgaggt
     6481 cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc gagctgctca ccacccactc
     6541 gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccgatc
     6601 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     6661 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     6721 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6781 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat
     6841 tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     6901 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6961 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     7021 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     7081 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7141 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7201 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7261 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7321 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7381 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7441 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7501 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7561 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7621 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7681 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7741 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7801 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7861 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7921 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7981 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8041 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8101 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8161 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8221 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8281 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8341 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8401 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8461 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8521 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8581 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8641 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8701 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8761 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8821 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8881 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8941 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9001 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     9061 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     9121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     9181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     9241 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     9301 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     9361 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     9421 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     9481 ttt
//
