LOCUS       V031679                 8202 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031679
VERSION     V031679
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..3008
                     /label="Phactr4(NM_175306)"
                     /note="Phactr4(NM_175306)"
                     /gene="Phactr4"
     CDS             3045..3764
                     /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    3802..4026
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4072..4500
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4514..4843
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4910..5704
                     /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    5878..5999
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6048..6064)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6072..6088
                     /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(6096..6126)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6141..6162
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6450..7035)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7206..8066)
                     /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(8067..8171)
                     /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 gccaccatgg aagacccatc ggaagaagca gagcagccct
      961 ctggagaccc aggcatgggc atggacagtg tggaagcagg ggacacaaca cctcccacca
     1021 aaaggaaaag caagttctca gcgttaggca agatcttcaa gccgtggaaa tggaggaaaa
     1081 aaaaaagcag tgataagttc aaagagacat cagaagtttt agagcgaaaa atatctatgc
     1141 gaaaaccaag agaagaactg gttaaacgag gggttctgtt ggaggacccc gagcaggatg
     1201 gtgaggattc aggaaagctg agccatgctg cattaaagaa cggacacacc acccccatag
     1261 gcagtgccag gtcctctagc ccagtcctgg tagaagaaga gccagagaga agtctcagga
     1321 acctcactcc agaagaggag tcaaagaaga gacttggctc aactggaagc cagcctaact
     1381 ctgaagccga gcctggtcct gagcatgcac ctaagcagcc tctgctgccc cccaagagac
     1441 ccttgtcctc ttcttgtgaa gcaaaggagg ttccggctgg cagtacagca aggtccgtct
     1501 cctccacctc tggctctacc accgtgacat ctgctgccac cactgctgcc acagacatga
     1561 caaagactgt taagtccttt gtgggcccca cccccgctcc tgcccccgcc cccaggactc
     1621 tgcctgctgc tcctgccagt gctaacactg ctgccaccac tacagcccct gccaagcaac
     1681 cccccatccc accccctaaa ccagctcaga gaaacagcaa ccccatcatt gctgaactat
     1741 cccaagcaat gaacagtggc acagtgctgt cgaaaccatc cccacccttg ccacccaaga
     1801 gaggcattcc atcaacttcg atacccagct tggagcctgc tgcttcgttc acaactaaga
     1861 cagcaaatga tcaaagagaa aaaactgtct ctttgtgttt ggaaccacca ctgataatcc
     1921 cgccttcgtc cccatcaccc ccactgccta cccacatacc tcccgagcct ccacggtccc
     1981 ctctagtgcc tgctaagact tttcaaattg tgccagaggt tgagttttcc tcttcttcag
     2041 atctattcca ggacatttcc cagcaggagg atcagaaaac agaagtcccc aagaagatac
     2101 aggatcagag ctttggggag tcccatatac cctccaggct gcccccactc ccactgcata
     2161 tccgaattca gcaggccctt accagccccc ttcctgtgac ccctcccttg gaaggcactc
     2221 acagagccca ttcactgctt tttgagaaca gtgacagctt ctccgaggac actggtaccc
     2281 tgggtcgtac caggtcactt cccatcacga ttgagatgtt gaaagttcca gatgatgaag
     2341 aggaagagca gacctgcccg tttgttgaag acgtgacgtc tacctcagcc actcctagcc
     2401 taccactgtg tctgcgagag gaagagaagg agagtgactc agattcagag ggtcccatta
     2461 agtaccgaga tgaagaagaa gatgatgatg atgatgaaag ccatcagagt gccctggcca
     2521 acagagtgaa gaggaaggac acgctggcaa tgaagctgag cagcagaccc agtgaaccag
     2581 agaccaacct gaattcttgg cctcgtaaaa gcaaggagga gtggaatgaa atacggcacc
     2641 agatcgggaa cacgctgatc cggcggctga gtcagagacc aacagcagaa gaattggagc
     2701 agcggaacat actacagcct aaaaatgaag ctgatcggca agcagagaaa cgggaaatca
     2761 agcgtcgcct cactagaaag ctcagtcaaa ggccaactgt ggctgaactg cttgccagga
     2821 agattctgcg gtttaatgag tatgtggaag taactgatgc tcatgactat gaccggcgag
     2881 cagacaaacc atggaccaaa ctgactcctg ctgacaaggc tgccataagg aaagaattaa
     2941 atgaatttaa gagctcggag atggaggttc atgtagacag caaacacttt acacgctacc
     3001 atcgcccatc gagtggcggt ggaggaagcg gcggaggtgg atctatggtg agcaagggcg
     3061 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3121 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3181 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3241 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3301 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3361 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3421 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3481 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3541 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3601 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3661 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3721 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtagtctaga gggcccgttt
     3781 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3841 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3901 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3961 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4021 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     4081 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4141 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4201 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4261 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4321 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4381 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4441 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4501 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4561 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4621 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4681 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4741 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4801 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4861 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4921 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4981 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5041 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     5101 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5161 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5221 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5281 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5341 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5401 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5461 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5521 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5581 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5641 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5701 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5761 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5821 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5881 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5941 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6001 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6061 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6121 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6241 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6301 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6361 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6961 ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga
     7021 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7081 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7201 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7261 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7321 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7381 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7441 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7501 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7561 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7621 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7681 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7741 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7801 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7861 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7921 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7981 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8041 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8101 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8161 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
