LOCUS       V029524                 8113 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029524
VERSION     V029524
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..3603
                     /label="HSPA5(NM_005347)"
                     /note="HSPA5(NM_005347)"
                     /gene="HSPA5"
     polyA_signal    3680..3904
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3950..4378
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4392..4721
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4788..5582
                     /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    5756..5877
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5926..5942)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5950..5966
                     /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(5974..6004)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6019..6040
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6328..6913)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7084..7944)
                     /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(7945..8049)
                     /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 catgaagctc tccctggtgg ccgcgatgct gctgctgctc
     1681 agcgcggcgc gggccgagga ggaggacaag aaggaggacg tgggcacggt ggtcggcatc
     1741 gacctgggga ccacctactc ctgcgtcggc gtgttcaaga acggccgcgt ggagatcatc
     1801 gccaacgatc agggcaaccg catcacgccg tcctatgtcg ccttcactcc tgaaggggaa
     1861 cgtctgattg gcgatgccgc caagaaccag ctcacctcca accccgagaa cacggtcttt
     1921 gacgccaagc ggctcatcgg ccgcacgtgg aatgacccgt ctgtgcagca ggacatcaag
     1981 ttcttgccgt tcaaggtggt tgaaaagaaa actaaaccat acattcaagt tgatattgga
     2041 ggtgggcaaa caaagacatt tgctcctgaa gaaatttctg ccatggttct cactaaaatg
     2101 aaagaaaccg ctgaggctta tttgggaaag aaggttaccc atgcagttgt tactgtacca
     2161 gcctatttta atgatgccca acgccaagca accaaagacg ctggaactat tgctggccta
     2221 aatgttatga ggatcatcaa cgagcctacg gcagctgcta ttgcttatgg cctggataag
     2281 agggaggggg agaagaacat cctggtgttt gacctgggtg gcggaacctt cgatgtgtct
     2341 cttctcacca ttgacaatgg tgtcttcgaa gttgtggcca ctaatggaga tactcatctg
     2401 ggtggagaag actttgacca gcgtgtcatg gaacacttca tcaaactgta caaaaagaag
     2461 acgggcaaag atgtcaggaa agacaataga gctgtgcaga aactccggcg cgaggtagaa
     2521 aaggccaaac gggccctgtc ttctcagcat caagcaagaa ttgaaattga gtccttctat
     2581 gaaggagaag acttttctga gaccctgact cgggccaaat ttgaagagct caacatggat
     2641 ctgttccggt ctactatgaa gcccgtccag aaagtgttgg aagattctga tttgaagaag
     2701 tctgatattg atgaaattgt tcttgttggt ggctcgactc gaattccaaa gattcagcaa
     2761 ctggttaaag agttcttcaa tggcaaggaa ccatcccgtg gcataaaccc agatgaagct
     2821 gtagcgtatg gtgctgctgt ccaggctggt gtgctctctg gtgatcaaga tacaggtgac
     2881 ctggtactgc ttgatgtatg tccccttaca cttggtattg aaactgtggg aggtgtcatg
     2941 accaaactga ttccaaggaa cacagtggtg cctaccaaga agtctcagat cttttctaca
     3001 gcttctgata atcaaccaac tgttacaatc aaggtctatg aaggtgaaag acccctgaca
     3061 aaagacaatc atcttctggg tacatttgat ctgactggaa ttcctcctgc tcctcgtggg
     3121 gtcccacaga ttgaagtcac ctttgagata gatgtgaatg gtattcttcg agtgacagct
     3181 gaagacaagg gtacagggaa caaaaataag atcacaatca ccaatgacca gaatcgcctg
     3241 acacctgaag aaatcgaaag gatggttaat gatgctgaga agtttgctga ggaagacaaa
     3301 aagctcaagg agcgcattga tactagaaat gagttggaaa gctatgccta ttctctaaag
     3361 aatcagattg gagataaaga aaagctggga ggtaaacttt cctctgaaga taaggagacc
     3421 atggaaaaag ctgtagaaga aaagattgaa tggctggaaa gccaccaaga tgctgacatt
     3481 gaagacttca aagctaagaa gaaggaactg gaagaaattg ttcaaccaat tatcagcaaa
     3541 ctctatggaa gtgcaggccc tcccccaact ggtgaagagg atacagcaga aaaagatgag
     3601 ttgtagggat cctgatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa
     3661 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3721 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3781 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3841 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3901 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4261 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4321 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4381 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4441 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4561 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4621 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4681 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4741 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4801 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4861 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4921 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4981 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5041 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5101 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5161 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5221 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5281 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5341 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5401 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5461 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5521 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5581 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5641 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5701 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5761 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5821 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5881 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5941 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6001 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6061 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6121 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6841 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6901 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6961 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7021 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     7081 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     7141 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     7201 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     7261 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     7321 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     7381 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     7441 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     7501 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     7561 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7621 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7681 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7741 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7801 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7861 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7921 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7981 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     8041 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     8101 gatcccctat ggt
//
