LOCUS       V026579                 8143 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026579
VERSION     V026579
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..3633
                     /label="SDHA(NM_004168)"
                     /note="SDHA(NM_004168)"
                     /gene="SDHA"
     polyA_signal    3710..3934
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3980..4408
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4422..4751
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4818..5612
                     /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    5786..5907
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5956..5972)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5980..5996
                     /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(6004..6034)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6049..6070
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6358..6943)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7114..7974)
                     /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(7975..8079)
                     /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 catgtcgggg gtccggggcc tgtcgcggct gctgagcgct
     1681 cggcgcctgg cgctggccaa ggcgtggcca acagtgttgc aaacaggaac ccgaggtttt
     1741 cacttcactg ttgatgggaa caagagggca tctgctaaag tttcagattc catttctgct
     1801 cagtatccag tagtggatca tgaatttgat gcagtggtgg taggcgctgg aggggcaggc
     1861 ttgcgagctg catttggcct ttctgaggca gggtttaata cagcatgtgt taccaagctg
     1921 tttcctacca ggtcacacac tgttgcagca cagggaggaa tcaatgctgc tctggggaac
     1981 atggaggagg acaactggag gtggcatttc tacgacaccg tgaagggctc cgactggctg
     2041 ggggaccagg atgccatcca ctacatgacg gagcaggccc ccgccgccgt ggtcgagcta
     2101 gaaaattatg gcatgccgtt tagcagaact gaagatggga agatttatca gcgtgcattt
     2161 ggtggacaga gcctcaagtt tggaaagggc gggcaggccc atcggtgctg ctgtgtggct
     2221 gatcggactg gccactcgct attgcacacc ttatatggaa ggtctctgcg atatgatacc
     2281 agctattttg tggagtattt tgccttggat ctcctgatgg agaatgggga gtgccgtggt
     2341 gtcatcgcac tgtgcataga ggacgggtcc atccatcgca taagagcaaa gaacactgtt
     2401 gttgccacag gaggctacgg gcgcacctac ttcagctgca cgtctgccca caccagcact
     2461 ggcgacggca cggccatgat caccagggca ggccttcctt gccaggacct agagtttgtt
     2521 cagttccacc ctacaggcat atatggtgct ggttgtctca ttacggaagg atgtcgtgga
     2581 gagggaggca ttctcattaa cagtcaaggc gaaaggttta tggagcgata cgcccctgtc
     2641 gcgaaggacc tggcgtctag agatgtggtg tctcggtcca tgactctgga gatccgagaa
     2701 ggaagaggct gtggccctga gaaagatcac gtctacctgc agctgcacca cctacctcca
     2761 gagcagctgg ccacgcgcct gcctggcatt tcagagacag ccatgatctt cgctggcgtg
     2821 gacgtcacga aggagccgat ccctgtcctc cccaccgtgc attataacat gggcggcatt
     2881 cccaccaact acaaggggca ggtcctgagg cacgtgaatg gccaggatca gattgtgccc
     2941 ggcctgtacg cctgtgggga ggccgcctgt gcctcggtac atggtgccaa ccgcctcggg
     3001 gcaaactcgc tcttggacct ggttgtcttt ggtcgggcat gtgccctgag catcgaagag
     3061 tcatgcaggc ctggagataa agtccctcca attaaaccaa acgctgggga agaatctgtc
     3121 atgaatcttg acaaattgag atttgctgat ggaagcataa gaacatcgga actgcgactc
     3181 agcatgcaga agtcaatgca aaatcatgct gccgtgttcc gtgtgggaag cgtgttgcaa
     3241 gaaggttgtg ggaaaatcag caagctctat ggagacctaa agcacctgaa gacgttcgac
     3301 cggggaatgg tctggaacac ggacctggtg gagaccctgg agctgcagaa cctgatgctg
     3361 tgtgcgctgc agaccatcta cggagcagag gcacggaagg agtcacgggg cgcgcatgcc
     3421 agggaagact acaaggtgcg gattgatgag tacgattact ccaagcccat ccaggggcaa
     3481 cagaagaagc cctttgagga gcactggagg aagcacaccc tgtcctatgt ggacgttggc
     3541 actgggaagg tcactctgga atatagaccc gtgatcgaca aaactttgaa cgaggctgac
     3601 tgtgccaccg tcccgccagc cattcgctcc tactgaggat cctgatatcc agcacagtgg
     3661 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3721 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3781 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3841 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3901 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3961 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4021 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4081 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4141 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4201 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4261 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4321 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4381 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     4441 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4501 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4621 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4681 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4741 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4801 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4861 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4921 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4981 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     5041 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     5101 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     5161 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5221 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5281 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5341 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5401 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5461 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5521 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     5581 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     5641 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5701 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5761 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5821 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5881 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5941 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6001 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6061 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6121 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6181 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6241 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6301 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6361 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6421 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6481 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6541 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6601 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6661 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6721 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6781 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6841 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6961 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7021 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7081 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7141 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7201 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7261 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7321 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7381 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7441 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7501 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7561 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7621 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7681 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7741 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7801 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7861 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7921 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7981 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8041 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8101 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
