LOCUS       V018370                 8348 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018370
VERSION     V018370
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"
     CDS             665..3028
                     /label="ITGB3(NM_000212)"
                     /note="ITGB3(NM_000212)"
                     /gene="ITGB3"
     CDS             3035..3100
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3124..3712
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3778..3826
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4028..4456
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4470..4799
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4866..5660
                     /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    5836..5957
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6006..6022)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6030..6046
                     /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(6054..6084)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6099..6120
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6408..6996)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7167..8027)
                     /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(8028..8132)
                     /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 gctctggcta gcgtttaaac ttaagcttgc
      661 caccatgcga gcgcggccgc ggccccggcc gctctgggcg actgtgctgg cgctgggggc
      721 gctggcgggc gttggcgtag gagggcccaa catctgtacc acgcgaggtg tgagctcctg
      781 ccagcagtgc ctggctgtga gccccatgtg tgcctggtgc tctgatgagg ccctgcctct
      841 gggctcacct cgctgtgacc tgaaggagaa tctgctgaag gataactgtg ccccagaatc
      901 catcgagttc ccagtgagtg aggcccgagt actagaggac aggcccctca gcgacaaggg
      961 ctctggagac agctcccagg tcactcaagt cagtccccag aggattgcac tccggctccg
     1021 gccagatgat tcgaagaatt tctccatcca agtgcggcag gtggaggatt accctgtgga
     1081 catctactac ttgatggacc tgtcttactc catgaaggat gatctgtgga gcatccagaa
     1141 cctgggtacc aagctggcca cccagatgcg aaagctcacc agtaacctgc ggattggctt
     1201 cggggcattt gtggacaagc ctgtgtcacc atacatgtat atctccccac cagaggccct
     1261 cgaaaacccc tgctatgata tgaagaccac ctgcttgccc atgtttggct acaaacacgt
     1321 gctgacgcta actgaccagg tgacccgctt caatgaggaa gtgaagaagc agagtgtgtc
     1381 acggaaccga gatgccccag agggtggctt tgatgccatc atgcaggcta cagtctgtga
     1441 tgaaaagatt ggctggagga atgatgcatc ccacttgctg gtgtttacca ctgatgccaa
     1501 gactcatata gcattggacg gaaggctggc aggcattgtc cagcctaatg acgggcagtg
     1561 tcatgttggt agtgacaatc attactctgc ctccactacc atggattatc cctctttggg
     1621 gctgatgact gagaagctat cccagaaaaa catcaatttg atctttgcag tgactgaaaa
     1681 tgtagtcaat ctctatcaga actatagtga gctcatccca gggaccacag ttggggttct
     1741 gtccatggat tccagcaatg tcctccagct cattgttgat gcttatggga aaatccgttc
     1801 taaagtagag ctggaagtgc gtgacctccc tgaagagttg tctctatcct tcaatgccac
     1861 ctgcctcaac aatgaggtca tccctggcct caagtcttgt atgggactca agattggaga
     1921 cacggtgagc ttcagcattg aggccaaggt gcgaggctgt ccccaggaga aggagaagtc
     1981 ctttaccata aagcccgtgg gcttcaagga cagcctgatc gtccaggtca cctttgattg
     2041 tgactgtgcc tgccaggccc aagctgaacc taatagccat cgctgcaaca atggcaatgg
     2101 gacctttgag tgtggggtat gccgttgtgg gcctggctgg ctgggatccc agtgtgagtg
     2161 ctcagaggag gactatcgcc cttcccagca ggacgaatgc agcccccggg agggtcagcc
     2221 cgtctgcagc cagcggggcg agtgcctctg tggtcaatgt gtctgccaca gcagtgactt
     2281 tggcaagatc acgggcaagt actgcgagtg tgacgacttc tcctgtgtcc gctacaaggg
     2341 ggagatgtgc tcaggccatg gccagtgcag ctgtggggac tgcctgtgtg actccgactg
     2401 gaccggctac tactgcaact gtaccacgcg tactgacacc tgcatgtcca gcaatgggct
     2461 gctgtgcagc ggccgcggca agtgtgaatg tggcagctgt gtctgtatcc agccgggctc
     2521 ctatggggac acctgtgaga agtgccccac ctgcccagat gcctgcacct ttaagaaaga
     2581 atgtgtggag tgtaagaagt ttgaccgggg agccctacat gacgaaaata cctgcaaccg
     2641 ttactgccgt gacgagattg agtcagtgaa agagcttaag gacactggca aggatgcagt
     2701 gaattgtacc tataagaatg aggatgactg tgtcgtcaga ttccagtact atgaagattc
     2761 tagtggaaag tccatcctgt atgtggtaga agagccagag tgtcccaagg gccctgacat
     2821 cctggtggtc ctgctctcag tgatgggggc cattctgctc attggccttg ccgccctgct
     2881 catctggaaa ctcctcatca ccatccacga ccgaaaagaa ttcgctaaat ttgaggaaga
     2941 acgcgccaga gcaaaatggg acacagccaa caacccactg tataaagagg ccacgtctac
     3001 cttcaccaat atcacgtacc ggggcactct cgaggactac aaagaccatg acggtgatta
     3061 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc
     3121 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3181 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3241 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3301 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3361 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3421 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3481 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3541 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3601 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3661 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     3721 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3781 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3841 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3901 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3961 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4021 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4081 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4141 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4201 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4261 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4321 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4381 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4441 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4501 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4561 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4621 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4681 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4741 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4801 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4861 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4921 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4981 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5041 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5101 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5161 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5221 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5281 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5341 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5401 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5461 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5521 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5581 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5641 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5701 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5761 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5821 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5881 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5941 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6001 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6061 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6121 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6601 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6841 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8161 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8221 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8281 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8341 tgcttagg
//
