LOCUS       V020135                 8546 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020135
VERSION     V020135
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..4036
                     /label="ITGA4(NM_000885)"
                     /note="ITGA4(NM_000885)"
                     /gene="ITGA4"
     CDS             4043..4108
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4146..4370
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4416..4844
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4858..5187
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5254..6048
                     /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    6222..6343
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6392..6408)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6416..6432
                     /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(6440..6470)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6485..6506
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6794..7379)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7550..8410)
                     /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(8411..8515)
                     /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 atggcttggg aagcgaggcg
      961 cgaacccggc ccccgaaggg ccgccgtccg ggagacggtg atgctgttgc tgtgcctggg
     1021 ggtcccgacc ggccgcccct acaacgtgga cactgagagc gcgctgcttt accagggccc
     1081 ccacaacacg ctgttcggct actcggtcgt gctgcacagc cacggggcga accgatggct
     1141 cctagtgggt gcgcccactg ccaactggct cgccaacgct tcagtgatca atcccggggc
     1201 gatttacaga tgcaggatcg gaaagaatcc cggccagacg tgcgaacagc tccagctggg
     1261 tagccctaat ggagaacctt gtggaaagac ttgtttggaa gagagagaca atcagtggtt
     1321 gggggtcaca ctttccagac agccaggaga aaatggatcc atcgtgactt gtgggcatag
     1381 atggaaaaat atattttaca taaagaatga aaataagctc cccactggtg gttgctatgg
     1441 agtgccccct gatttacgaa cagaactgag taaaagaata gctccgtgtt atcaagatta
     1501 tgtgaaaaaa tttggagaaa attttgcatc atgtcaagct ggaatatcca gtttttacac
     1561 aaaggattta attgtgatgg gggccccagg atcatcttac tggactggct ctctttttgt
     1621 ctacaatata actacaaata aatacaaggc ttttttagac aaacaaaatc aagtaaaatt
     1681 tggaagttat ttaggatatt cagtcggagc tggtcatttt cggagccagc atactaccga
     1741 agtagtcgga ggagctcctc aacatgagca gattggtaag gcatatatat tcagcattga
     1801 tgaaaaagaa ctaaatatct tacatgaaat gaaaggtaaa aagcttggat cgtactttgg
     1861 agcttctgtc tgtgctgtgg acctcaatgc agatggcttc tcagatctgc tcgtgggagc
     1921 acccatgcag agcaccatca gagaggaagg aagagtgttt gtgtacatca actctggctc
     1981 gggagcagta atgaatgcaa tggaaacaaa cctcgttgga agtgacaaat atgctgcaag
     2041 atttggggaa tctatagtta atcttggcga cattgacaat gatggctttg aagatgttgc
     2101 tatcggagct ccacaagaag atgacttgca aggtgctatt tatatttaca atggccgtgc
     2161 agatgggatc tcgtcaacct tctcacagag aattgaagga cttcagatca gcaaatcgtt
     2221 aagtatgttt ggacagtcta tatcaggaca aattgatgca gataataatg gctatgtaga
     2281 tgtagcagtt ggtgcttttc ggtctgattc tgctgtcttg ctaaggacaa gacctgtagt
     2341 aattgttgac gcttctttaa gccaccctga gtcagtaaat agaacgaaat ttgactgtgt
     2401 tgaaaatgga tggccttctg tgtgcataga tctaacactt tgtttctcat ataagggcaa
     2461 ggaagttcca ggttacattg ttttgtttta taacatgagt ttggatgtga acagaaaggc
     2521 agagtctcca ccaagattct atttctcttc taatggaact tctgacgtga ttacaggaag
     2581 catacaggtg tccagcagag aagctaactg tagaacacat caagcattta tgcggaaaga
     2641 tgtgcgggac atcctcaccc caattcagat tgaagctgct taccaccttg gtcctcatgt
     2701 catcagtaaa cgaagtacag aggaattccc accacttcag ccaattcttc agcagaagaa
     2761 agaaaaagac ataatgaaaa aaacaataaa ctttgcaagg ttttgtgccc atgaaaattg
     2821 ttctgctgat ttacaggttt ctgcaaagat tgggtttttg aagccccatg aaaataaaac
     2881 atatcttgct gttgggagta tgaagacatt gatgttgaat gtgtccttgt ttaatgctgg
     2941 agatgatgca tatgaaacga ctctacatgt caaactaccc gtgggtcttt atttcattaa
     3001 gattttagag ctggaagaga agcaaataaa ctgtgaagtc acagataact ctggcgtggt
     3061 acaacttgac tgcagtattg gctatatata tgtagatcat ctctcaagga tagatattag
     3121 ctttctcctg gatgtgagct cactcagcag agcggaagag gacctcagta tcacagtgca
     3181 tgctacctgt gaaaatgaag aggaaatgga caatctaaag cacagcagag tgactgtagc
     3241 aataccttta aaatatgagg ttaagctgac tgttcatggg tttgtaaacc caacttcatt
     3301 tgtgtatgga tcaaatgatg aaaatgagcc tgaaacgtgc atggtggaga aaatgaactt
     3361 aactttccat gttatcaaca ctggcaatag tatggctccc aatgttagtg tggaaataat
     3421 ggtaccaaat tcttttagcc cccaaactga taagctgttc aacattttgg atgtccagac
     3481 tactactgga gaatgccact ttgaaaatta tcaaagagtg tgtgcattag agcagcaaaa
     3541 gagtgcaatg cagaccttga aaggcatagt ccggttcttg tccaagactg ataagaggct
     3601 attgtactgc ataaaagctg atccacattg tttaaatttc ttgtgtaatt ttgggaaaat
     3661 ggaaagtgga aaagaagcca gtgttcatat ccaactggaa ggccggccat ccattttaga
     3721 aatggatgag acttcagcac tcaagtttga aataagagca acaggttttc cagagccaaa
     3781 tccaagagta attgaactaa acaaggatga gaatgttgcg catgttctac tggaaggact
     3841 acatcatcaa agacccaaac gttatttcac catagtgatt atttcaagta gcttgctact
     3901 tggacttatt gtacttctat tgatctcata tgttatgtgg aaggctggct tctttaaaag
     3961 acaatacaaa tctatcctac aagaagaaaa cagaagagac agttggagtt atatcaacag
     4021 taaaagcaat gatgatctcg aggactacaa agaccatgac ggtgattata aagatcatga
     4081 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     4141 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4201 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4261 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4321 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     4381 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     4441 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4501 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4561 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4621 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4681 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4741 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4801 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4861 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4921 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5041 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5101 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     5161 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5221 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5281 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5341 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5401 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5461 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5521 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5581 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5641 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5701 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5761 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5821 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5881 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5941 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6001 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6061 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6121 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6181 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6361 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7441 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7501 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7561 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7621 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7681 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7741 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7801 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7861 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7921 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7981 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8041 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8101 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8161 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8221 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8281 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8341 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8401 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8461 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8521 ttccccgaaa agtgccacct gacgtc
//
