LOCUS       V035595                10405 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035595
VERSION     V035595
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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"
     CDS             1642..5895
                     /label="TNS2(NM_001416204)"
                     /note="TNS2(NM_001416204)"
                     /gene="TNS2"
     polyA_signal    5972..6196
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      6242..6670
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6684..7013
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             7080..7874
                     /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    8048..8169
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8218..8234)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8242..8258
                     /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(8266..8296)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8311..8332
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8620..9205)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9376..10236)
                     /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(10237..10341)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc
     1621 ggaggcggag ggagtgaatt catgaagtcc agcggccctg tggagaggct gctcagagcc
     1681 ctggggagga gggacagcag ccgggccgca agcaggccta ggaaagctga gcctcatagc
     1741 ttccgggaga aggttttccg gaagaaacct ccagtctgtg cagtatgtaa ggtgaccatc
     1801 gatgggacag gcgtttcgtg cagagtctgc aaggtggcga cgcacagaaa atgtgaagca
     1861 aaggtgactt cagcctgtca ggccttgcct cccgtggagt tgcggcgaaa cacggcccca
     1921 gtcaggcgca tagagcacct gggatccacc aaatctctga accactcaaa gcagcgcagc
     1981 actctgccca ggagcttcag cctggacccg ctcatggagc ggcgctggga cttagacctc
     2041 acctacgtga cggagcgcat cttggccgcc gccttccccg cgcggcccga tgaacagcgg
     2101 caccggggcc acctgcgcga gctggcccat gtgctgcaat ccaagcaccg ggacaagtac
     2161 ctgctcttca acctttcaga gaaaaggcat gacctgaccc gcttaaaccc caaggttcaa
     2221 gacttcggct ggcctgagct gcatgctcca cccctggaca agctgtgctc catctgcaaa
     2281 gccatggaga catggctcag tgctgaccca cagcacgtgg tcgtactata ctgcaaggga
     2341 aacaagggca agcttggggt catcgtttct gcctacatgc actacagcaa gatctctgca
     2401 ggggcggacc aggcactggc cactcttacc atgcggaaat tctgcgagga caaggtggcc
     2461 acagaactgc agccctccca gcgtcgatat atcagctact tcagtgggct gctatctggc
     2521 tccatcagaa tgaacagcag ccctctcttc ctgcactatg tgctcatccc catgctgcca
     2581 gcctttgaac ctggcacagg cttccagccc ttccttaaaa tctaccagtc catgcagctt
     2641 gtctacacat ctggagtcta tcacattgca ggccctggtc cccagcagct ttgcatcagc
     2701 ctggagccag ccctcctcct caaaggcgat gtcatggtaa catgttatca caagggtggc
     2761 cggggcacag accggaccct cgtgttccga gtccagttcc acacctgcac catccacgga
     2821 ccacagctca ctttccccaa ggaccagctt gacgaggcct ggactgatga gaggttcccc
     2881 ttccaagcct ccgtggagtt tgtcttctcc tccagccccg agaagatcaa aggcagcact
     2941 ccacggaacg acccctcggt ctctgtcgac tacaacacca ctgagccagc cgtgcgctgg
     3001 gactcctatg agaacttcaa ccagcaccac gaggacagtg tggatggtgc gcaggcagcc
     3061 tgcagggtgg gaggctcctt gacccacacc cggggtcccc tggatggcag tccttatgcc
     3121 caggtgcagc ggcctccccg gcagaccccc ccggcaccct ctccagagcc tccaccaccc
     3181 cccatgctct ctgtcagcag cgactcaggc cattcctcca cgctgaccac agagccggct
     3241 gctgagtccc ctggccggcc gccccctaca gctgctgaac ggcaggagct ggatcgcctc
     3301 ctaggaggct gcggagtggc cagtgggggc cggggagctg ggcgcgagac ggccatccta
     3361 gatgacgaag agcagcccac tgtgggcgga ggcccccacc tcggagtgta tccaggccat
     3421 aggcctggcc tcagccgcca ctgctcctgc cgccagggct accgggagcc ctgcggggtt
     3481 cccaatgggg gctactaccg gccagaggga accctggaga ggaggcgact ggcctacggg
     3541 ggctatgagg gatcccccca gggctacgcc gaggcctcga tggagaagag gcgcctctgc
     3601 cgatcgctgt cagaggggct atacccctac ccacctgaga tggggaaacc agccactggg
     3661 gactttggct accgcgcccc aggctaccgg gaggtggtca tcctggagga ccctgggctg
     3721 cctgccctat acccatgccc agcctgcgag gagaagctgg cgctgcctac agcagccttg
     3781 tatggactgc ggctggagag ggaggctgga gaagggtggg caagtgaggc tggcaagcct
     3841 ctcctgcacc cagtgcggcc tgggcacccg ctgcctctgc tcttgcctgc ctgtgggcat
     3901 caccatgccc cgatgcctga ctacagctgc ctgaagccac ccaaggcagg cgaggaaggg
     3961 cacgagggct gctcctacac catgtgcccc gaaggcaggt atgggcatcc agggtaccct
     4021 gccctggtga catacagcta tggaggagca gttcccagtt actgcccagc atatggccgt
     4081 gtgcctcata gctgtggctc tccaggagag ggcagagggt atcccagccc tggtgcccac
     4141 tccccacggg ctggctccat ttccccgggc agcccgccct atccacaatc taggaagctg
     4201 agctacgaga tccctacgga ggagggaggg gacaggtacc cattgcctgg gcacctggcc
     4261 tcagcaggac ctttggcatc tgcagagtcg ctggagccgg tgtcctggag ggagggcccc
     4321 agtgggcaca gcacactgcc tcggtctccc cgagatgccc catgcagtgc ttcgtcagag
     4381 ttgtctggtc cctccacgcc cctgcacacc agcagtccag tccagggcaa ggaaagcacc
     4441 cggcgacagg acaccaggtc ccccacctca gcgcccactc agagactgag tcctggcgag
     4501 gccttgcccc ctgtttccca ggcaggcacc ggaaaggccc ctgagctgcc gtcgggaagt
     4561 gggcctgagc ctctggcccc tagcccagtc tctccgacct tccctcccag ctcgcccagt
     4621 gactggcctc aggaaaggag tccagggggc cactcagatg gcgccagtcc tcggagccct
     4681 gtgcccacca cacttcctgg cctccgccac gccccctggc aaggccctcg aggccccccc
     4741 gacagcccag atgggtctcc cctcactcct gtgccttccc agatgccctg gcttgtggcc
     4801 agcccagagc cgcctcagag ctcacctaca cctgctttcc ccctggctgc ctcctatgac
     4861 accaatggcc ttagccagcc cccacttcct gagaaacgcc acctgcccgg gccggggcaa
     4921 cagccaggac cctggggccc agagcaggca tcatcgccag ccagaggcat cagtcaccat
     4981 gtcaccttcg cacctctgct ctcagataat gtcccccaaa ccccagagcc tcctacacaa
     5041 gagagccaaa gcaatgtcaa gtttgtccag gatacatcca agttctggta caagccacac
     5101 ctgtcccgtg accaagccat tgccctgctg aaggacaagg accctggggc cttcctgatc
     5161 agggacagtc attcattcca aggagcttat gggctggccc tcaaggtggc cacaccgcca
     5221 cccagtgccc agccctggaa aggggacccc gtggaacagc tggtccgcca tttcctcatc
     5281 gagactgggc ccaaaggggt gaagatcaag ggctgcccca gtgagcccta ctttggcagc
     5341 ctgtccgcct tggtctccca gcactccatc tcccccatct ccctgccctg ctgcctgcgc
     5401 attcccagca aagatcctct ggaagagacc ccagaggctc cagtgcccac caacatgagc
     5461 acagcggcag acctcctgcg tcagggtgct gcctgcagcg tgctctactt gacctcagtg
     5521 gagacagagt cactgacggg cccccaagct gtggcccggg ccagctctgc agctctgagc
     5581 tgtagccccc gcccgacacc agctgttgtc cacttcaagg tgtcagccca gggcattaca
     5641 ctgacggaca accaaaggaa gctcttcttt cgccgccatt atccagtgaa cagcatcacc
     5701 ttctccagca ctgaccctca agaccggaga tggaccaacc cagacgggac cacctccaag
     5761 atctttggtt tcgtggccaa gaagccggga agcccctggg agaatgtgtg tcacctcttt
     5821 gcagagcttg acccagatca gcctgctggc gccattgtca ccttcatcac caaagttcta
     5881 ctgggccaga gaaaatgagg atcctgatat ccagcacagt ggcggccgct cgagtctaga
     5941 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     6001 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     6061 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     6121 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     6181 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     6241 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6661 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6841 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6901 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     6961 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     7021 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     7081 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     7141 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     7201 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     7261 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     7321 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     7381 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     7441 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     7501 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     7561 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     7621 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     7681 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     7741 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     7801 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     7861 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     7921 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     7981 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     8041 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8161 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     8221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     8281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     8341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     8401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     8461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     8521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     8821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     9001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     9061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     9121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca agcagcagat
     9181 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9241 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9301 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     9361 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     9421 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     9481 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9541 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9601 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9661 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9721 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9781 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9841 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9901 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9961 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10021 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    10081 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10141 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10201 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10261 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10321 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    10381 gggagatctc ccgatcccct atggt
//
