LOCUS       V034604                 8430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034604
VERSION     V034604
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             927..3233
                     /label="ITGB2(NM_000211)"
                     /note="ITGB2(NM_000211)"
                     /gene="ITGB2"
     CDS             3273..3992
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    4030..4254
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4300..4728
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4742..5071
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5138..5932
                     /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    6106..6227
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6276..6292)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6300..6316
                     /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(6324..6354)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6369..6390
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6678..7263)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7434..8294)
                     /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(8295..8399)
                     /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 gtttaaagcc actgggatcc gccaccatgc tgggcctgcg ccccccactg ctcgccctgg
      961 tggggctgct ctccctcggg tgcgtcctct ctcaggagtg cacgaagttc aaggtcagca
     1021 gctgccggga atgcatcgag tcggggcccg gctgcacctg gtgccagaag ctgaacttca
     1081 cagggccggg ggatcctgac tccattcgct gcgacacccg gccacagctg ctcatgaggg
     1141 gctgtgcggc tgacgacatc atggacccca caagcctcgc tgaaacccag gaagaccaca
     1201 atgggggcca gaagcagctg tccccacaaa aagtgacgct ttacctgcga ccaggccagg
     1261 cagcagcgtt caacgtgacc ttccggcggg ccaagggcta ccccatcgac ctgtactatc
     1321 tgatggacct ctcctactcc atgcttgatg acctcaggaa tgtcaagaag ctaggtggcg
     1381 acctgctccg ggccctcaac gagatcaccg agtccggccg cattggcttc gggtccttcg
     1441 tggacaagac cgtgctgccg ttcgtgaaca cgcaccctga taagctgcga aacccatgcc
     1501 ccaacaagga gaaagagtgc cagcccccgt ttgccttcag gcacgtgctg aagctgacca
     1561 acaactccaa ccagtttcag accgaggtcg ggaagcagct gatttccgga aacctggatg
     1621 cacccgaggg tgggctggac gccatgatgc aggtcgccgc ctgcccggag gaaatcggct
     1681 ggcgcaacgt cacgcggctg ctggtgtttg ccactgatga cggcttccat ttcgcgggcg
     1741 acgggaagct gggcgccatc ctgaccccca acgacggccg ctgtcacctg gaggacaact
     1801 tgtacaagag gagcaacgaa ttcgactacc catcggtggg ccagctggcg cacaagctgg
     1861 ctgaaaacaa catccagccc atcttcgcgg tgaccagtag gatggtgaag acctacgaga
     1921 aactcaccga gatcatcccc aagtcagccg tgggggagct gtctgaggac tccagcaatg
     1981 tggtccatct cattaagaat gcttacaata aactctcctc cagggtcttc ctggatcaca
     2041 acgccctccc cgacaccctg aaagtcacct acgactcctt ctgcagcaat ggagtgacgc
     2101 acaggaacca gcccagaggt gactgtgatg gcgtgcagat caatgtcccg atcaccttcc
     2161 aggtgaaggt cacggccaca gagtgcatcc aggagcagtc gtttgtcatc cgggcgctgg
     2221 gcttcacgga catagtgacc gtgcaggttc ttccccagtg tgagtgccgg tgccgggacc
     2281 agagcagaga ccgcagcctc tgccatggca agggcttctt ggagtgcggc atctgcaggt
     2341 gtgacactgg ctacattggg aaaaactgtg agtgccagac acagggccgg agcagccagg
     2401 agctggaagg aagctgccgg aaggacaaca actccatcat ctgctcaggg ctgggggact
     2461 gtgtctgcgg gcagtgcctg tgccacacca gcgacgtccc cggcaagctg atatacgggc
     2521 agtactgcga gtgtgacacc atcaactgtg agcgctacaa cggccaggtc tgcggcggcc
     2581 cggggagggg gctctgcttc tgcgggaagt gccgctgcca cccgggcttt gagggctcag
     2641 cgtgccagtg cgagaggacc actgagggct gcctgaaccc gcggcgtgtt gagtgtagtg
     2701 gtcgtggccg gtgccgctgc aacgtatgcg agtgccattc aggctaccag ctgcctctgt
     2761 gccaggagtg ccccggctgc ccctcaccct gtggcaagta catctcctgc gccgagtgcc
     2821 tgaagttcga aaagggcccc tttgggaaga actgcagcgc ggcgtgtccg ggcctgcagc
     2881 tgtcgaacaa ccccgtgaag ggcaggacct gcaaggagag ggactcagag ggctgctggg
     2941 tggcctacac gctggagcag caggacggga tggaccgcta cctcatctat gtggatgaga
     3001 gccgagagtg tgtggcaggc cccaacatcg ccgccatcgt cgggggcacc gtggcaggca
     3061 tcgtgctgat cggcattctc ctgctggtca tctggaaggc tctgatccac ctgagcgacc
     3121 tccgggagta caggcgcttt gagaaggaga agctcaagtc ccagtggaac aatgataatc
     3181 cccttttcaa gagcgccacc acgacggtca tgaaccccaa gtttgctgag agtctctcga
     3241 gtggcggtgg aggaagcggc ggaggtggat ctatggtgag caagggcgag gagctgttca
     3301 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     3361 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     3421 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     3481 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     3541 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     3601 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     3661 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     3721 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     3781 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     3841 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     3901 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3961 tcactctcgg catggacgag ctgtacaagt agtctagagg gcccgtttaa acccgctgat
     4021 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4081 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4141 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4201 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4261 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4321 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4381 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4441 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4501 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4561 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4621 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4681 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4741 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4801 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4861 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4921 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4981 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5041 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5101 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5161 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5221 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5281 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5341 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5401 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5461 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5521 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5581 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5641 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5701 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5761 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5821 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5881 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5941 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6001 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6061 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6121 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6181 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6241 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6301 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6361 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6481 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6541 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6601 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7201 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7441 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7501 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7561 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7621 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7681 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7741 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7801 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7861 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7921 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7981 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8041 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8101 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8161 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8221 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8281 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8401 acatttcccc gaaaagtgcc acctgacgtc
//
