LOCUS       V018558                 9167 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018558
VERSION     V018558
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             671..3814
                     /label="ITGAV(NM_002210)"
                     /note="ITGAV(NM_002210)"
                     /gene="ITGAV"
     CDS             3854..3919
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3943..4531
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4597..4645
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4847..5275
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5289..5618
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5685..6479
                     /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    6655..6776
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6825..6841)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6849..6865
                     /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(6873..6903)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6918..6939
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7227..7815)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7986..8846)
                     /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(8847..8951)
                     /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 ttaagcttgg
      661 taccgccacc atggcttttc cgccgcggcg acggctgcgc ctcggtcccc gcggcctccc
      721 gcttcttctc tcgggactcc tgctacctct gtgccgcgcc ttcaacctag acgtggacag
      781 tcctgccgag tactctggcc ccgagggaag ttacttcggc ttcgccgtgg atttcttcgt
      841 gcccagcgcg tcttcccgga tgtttcttct cgtgggagct cccaaagcaa acaccaccca
      901 gcctgggatt gtggaaggag ggcaggtcct caaatgtgac tggtcttcta cccgccggtg
      961 ccagccaatt gaatttgatg caacaggcaa tagagattat gccaaggatg atccattgga
     1021 atttaagtcc catcagtggt ttggagcatc tgtgaggtcg aaacaggata aaattttggc
     1081 ctgtgcccca ttgtaccatt ggagaactga gatgaaacag gagcgagagc ctgttggaac
     1141 atgctttctt caagatggaa caaagactgt tgagtatgct ccatgtagat cacaagatat
     1201 tgatgctgat ggacagggat tttgtcaagg aggattcagc attgatttta ctaaagctga
     1261 cagagtactt cttggtggtc ctggtagctt ttattggcaa ggtcagctta tttcggatca
     1321 agtggcagaa atcgtatcta aatacgaccc caatgtttac agcatcaagt ataataacca
     1381 attagcaact cggactgcac aagctatttt tgatgacagc tatttgggtt attctgtggc
     1441 tgtcggagat ttcaatggtg atggcataga tgactttgtt tcaggagttc caagagcagc
     1501 aaggactttg ggaatggttt atatttatga tgggaagaac atgtcctcct tatacaattt
     1561 tactggcgag cagatggctg catatttcgg attttctgta gctgccactg acattaatgg
     1621 agatgattat gcagatgtgt ttattggagc acctctcttc atggatcgtg gctctgatgg
     1681 caaactccaa gaggtggggc aggtctcagt gtctctacag agagcttcag gagacttcca
     1741 gacgacaaag ctgaatggat ttgaggtctt tgcacggttt ggcagtgcca tagctccttt
     1801 gggagatctg gaccaggatg gtttcaatga tattgcaatt gctgctccat atgggggtga
     1861 agataaaaaa ggaattgttt atatcttcaa tggaagatca acaggcttga acgcagtccc
     1921 atctcaaatc cttgaagggc agtgggctgc tcgaagcatg ccaccaagct ttggctattc
     1981 aatgaaagga gccacagata tagacaaaaa tggatatcca gacttaattg taggagcttt
     2041 tggtgtagat cgagctatct tatacagggc cagaccagtt atcactgtaa atgctggtct
     2101 tgaagtgtac cctagcattt taaatcaaga caataaaacc tgctcactgc ctggaacagc
     2161 tctcaaagtt tcctgtttta atgttaggtt ctgcttaaag gcagatggca aaggagtact
     2221 tcccaggaaa cttaatttcc aggtggaact tcttttggat aaactcaagc aaaagggagc
     2281 aattcgacga gcactgtttc tctacagcag gtccccaagt cactccaaga acatgactat
     2341 ttcaaggggg ggactgatgc agtgtgagga attgatagcg tatctgcggg atgaatctga
     2401 atttagagac aaactcactc caattactat ttttatggaa tatcggttgg attatagaac
     2461 agctgctgat acaacaggct tgcaacccat tcttaaccag ttcacgcctg ctaacattag
     2521 tcgacaggct cacattctac ttgactgtgg tgaagacaat gtctgtaaac ccaagctgga
     2581 agtttctgta gatagtgatc aaaagaagat ctatattggg gatgacaacc ctctgacatt
     2641 gattgttaag gctcagaatc aaggagaagg tgcctacgaa gctgagctca tcgtttccat
     2701 tccactgcag gctgatttca tcggggttgt ccgaaacaat gaagccttag caagactttc
     2761 ctgtgcattt aagacagaaa accaaactcg ccaggtggta tgtgaccttg gaaacccaat
     2821 gaaggctgga actcaactct tagctggtct tcgtttcagt gtgcaccagc agtcagagat
     2881 ggatacttct gtgaaatttg acttacaaat ccaaagctca aatctatttg acaaagtaag
     2941 cccagttgta tctcacaaag ttgatcttgc tgttttagct gcagttgaga taagaggagt
     3001 ctcgagtcct gatcatgtct ttcttccgat tccaaactgg gagcacaagg agaaccctga
     3061 gactgaagaa gatgttgggc cagttgttca gcacatctat gagctgagaa acaatggtcc
     3121 aagttcattc agcaaggcaa tgctccatct tcagtggcct tacaaatata ataataacac
     3181 tctgttgtat atccttcatt atgatattga tggaccaatg aactgcactt cagatatgga
     3241 gatcaaccct ttgagaatta agatctcatc tttgcaaaca actgaaaaga atgacacggt
     3301 tgccgggcaa ggtgagcggg accatctcat cactaagcgg gatcttgccc tcagtgaagg
     3361 agatattcac actttgggtt gtggagttgc tcagtgcttg aagattgtct gccaagttgg
     3421 gagattagac agaggaaaga gtgcaatctt gtacgtaaag tcattactgt ggactgagac
     3481 ttttatgaat aaagaaaatc agaatcattc ctattctctg aagtcgtctg cttcatttaa
     3541 tgtcatagag tttccttata agaatcttcc aattgaggat atcaccaact ccacattggt
     3601 taccactaat gtcacctggg gcattcagcc agcgcccatg cctgtgcctg tgtgggtgat
     3661 cattttagca gttctagcag gattgttgct actggctgtt ttggtatttg taatgtacag
     3721 gatgggcttt tttaaacggg tccggccacc tcaagaagaa caagaaaggg agcagcttca
     3781 acctcatgaa aatggtgaag gaaactcaga aactgaattc tgcagatatc cagcacagtg
     3841 gcggccgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3901 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     3961 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     4021 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     4081 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     4141 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4201 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4261 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4321 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4381 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4441 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4501 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     4561 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4621 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4681 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4741 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4801 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     4861 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4921 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4981 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     5041 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5101 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5161 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     5221 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5281 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5341 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5401 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5461 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5521 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5581 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5641 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5701 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5761 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5821 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5881 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5941 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6001 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6061 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6121 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6181 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     6241 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6301 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6361 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6421 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6481 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6541 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6601 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6661 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6721 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6781 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6841 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6901 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6961 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7801 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7921 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7981 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8041 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8101 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8161 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8221 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8281 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8341 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8401 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8461 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8521 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8581 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8641 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8701 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8761 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8821 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8881 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8941 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     9001 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     9061 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     9121 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
