LOCUS       V041243                 8524 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041243
VERSION     V041243
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..4014
                     /label="JADE2(NM_001289984)"
                     /note="JADE2(NM_001289984)"
                     /gene="JADE2"
     polyA_signal    4091..4315
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4361..4789
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4803..5132
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5199..5993
                     /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    6167..6288
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6337..6353)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6361..6377
                     /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(6385..6415)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6430..6451
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6739..7324)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7495..8355)
                     /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(8356..8460)
                     /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 catggaagag aagaggcgaa aatactccat cagcagtgac
     1681 aactctgaca ccactgacag tcatgcgaca tctacatccg catcaagatg ctccaaactg
     1741 cccagcagca ccaagtcggg ctggccccga cagaacgaaa agaagccctc cgaggttttc
     1801 cggacagact tgatcacagc catgaagatc ccggactcat accagctcag cccggatgac
     1861 tactacatcc tggcagaccc atggcgacag gaatgggaga aaggtgtgca ggtgcctgcc
     1921 ggggcagagg ccatcccaga gcccgtggtg aggatcctcc caccactgga aggcccccct
     1981 gcccaggcat ccccgagcag caccatgctt ggtgagggct cccagcctga ttggccaggg
     2041 ggcagccgct atgacttgga cgagattgat gcctactggc tggagctcat caactcggag
     2101 cttaaggaga tggagaggcc ggagctggac gagctgacat tagagcgtgt gctggaggag
     2161 ctggagaccc tgtgccacca gaatatggcc agggccattg agacgcagga ggggctgggc
     2221 atcgagtacg acgaggatgt tgtctgcgac gtgtgtcgct ctcctgaggg cgaggatggc
     2281 aacgagatgg tcttctgtga caagtgcaac gtctgtgtgc atcaggcatg ctacgggatc
     2341 ctcaaggtgc ccacgggcag ctggctgtgc cggacgtgtg ccctgggtgt ccagccaaag
     2401 tgcctgctct gccccaagcg aggaggagcc ttgaagccca ctagaagtgg gaccaagtgg
     2461 gtgcatgtca gctgtgccct atggattcct gaggtcagca tcggctgccc agagaagatg
     2521 gagcccatca ccaagatctc gcatatccca gccagccgct gggctctgtc ctgcagcctc
     2581 tgcaaggaat gcacaggcac ctgcatccag tgttccatgc cttcctgcgt cacagcgttc
     2641 catgtcacat gcgcctttga ccacggcctg gaaatgcgga ctatattagc agacaacgat
     2701 gaggtcaagt tcaagtcatt ctgccaggag cacagtgacg ggggcccacg taatgagccc
     2761 acatctgagc ccacggaacc cagccaggct ggcgaggacc tggaaaaggt gaccctgcgc
     2821 aagcagcggc tgcagcagct agaggaggac ttctacgagc tggtggagcc ggctgaggtg
     2881 gctgagcggc tggacctggc tgaggcactg gtcgacttca tctaccagta ctggaagctg
     2941 aagaggaaag ccaatgccaa ccagccgctg ctgaccccca agaccgacga ggtggacaac
     3001 ctggcccagc aggagcagga cgtcctctac cgccgcctga agctcttcac ccatctgcgg
     3061 caggacctag agagggttag aaatctgtgc tacatggtga caaggcgcga gagaacgaaa
     3121 cacgccatct gcaaactcca ggagcagata ttccacctgc agatgaaact tattgaacag
     3181 gatctgtgtc gagcaggcct gtccacctca ttccccatcg atggcacctt cttcaacagc
     3241 tggctggcac agtcggtgca gatcacagca gagaacatgg ccatgagcga gtggccactg
     3301 aacaatgggc accgcgagga ccctgctcca gggctgctgt cagaggaact gctgcaggac
     3361 gaggagacac tgctcagctt catgcgggac ccctcgctgc gacctggtga ccctgctagg
     3421 aaggcccgag gccgcacccg cctgcctgcc aagaagaaac caccaccacc accaccgcag
     3481 gacgggcctg gttcacggac gactccagac aaagccccca agaagacctg gggccaggat
     3541 gcaggcagtg gcaagggggg tcaagggcca cctaccagga agccaccacg tcggacatct
     3601 tctcacttgc cgtccagccc tgcagccggg gactgtccca tcctagccac ccctgaaagc
     3661 cccccgccac tggcccctga gaccccggac gaggcagcct cagtagctgc tgactcagat
     3721 gtccaagtgc ctggccctgc agcaagccct aagcctttgg gccggctccg gccaccccgc
     3781 gagagcaagg taacccggag attgccgggt gccaggcctg atgctgggat gggaccacct
     3841 tcagctgtgg ctgagaggcc caaggtcagc ctgcattttg acactgagac tgatggctac
     3901 ttctctgatg gggagatgag cgactcagat gtagaggccg aggacggtgg ggtgcagcgg
     3961 ggtccccggg aggcaggggc agaggaggtg gtccgcatgg gcgtactggc ctcctaagga
     4021 tcctgatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     4081 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     4141 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     4201 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     4261 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     4321 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4501 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4801 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4861 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4921 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4981 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5041 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5101 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5161 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5221 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5281 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5341 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5401 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5461 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5521 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5581 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5641 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5701 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5761 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5821 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5881 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5941 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6001 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6061 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6121 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6301 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6541 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6601 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7141 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     7201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7261 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8461 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     8521 tggt
//
