LOCUS       V032399                 8662 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032399
VERSION     V032399
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..4152
                     /label="TELO2(NM_016111)"
                     /note="TELO2(NM_016111)"
                     /gene="TELO2"
     polyA_signal    4229..4453
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4499..4927
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4941..5270
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5337..6131
                     /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    6305..6426
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6475..6491)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6499..6515
                     /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(6523..6553)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6568..6589
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6877..7462)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7633..8493)
                     /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(8494..8598)
                     /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 catggagcca gcaccctcag aggttcgact cgccgtccgg
     1681 gaagccattc atgccctctc gtcttcggag gatggcggcc acatcttctg caccctggag
     1741 tccctgaagc ggtatctcgg tgagatggag cctccagcgc tcccgaggga gaaggaggag
     1801 tttgcctcgg cccacttctc gcctgtcctc agatgtcttg ccagcaggct gagcccagcc
     1861 tggctggagc tgctgcccca tggccgcctg gaggagctgt gggccagctt cttcctggag
     1921 ggcccggcgg accaagcctt cctggtgttg atggagacca tcgagggtgc tgcgggcccc
     1981 agcttccggc tgatgaagat ggcgcggctg ctggccagat tcctgcgcga gggccggctg
     2041 gcagtgctga tggaggcgca gtgtcggcag cagacgcagc ccggcttcat cctgctccgg
     2101 gagacgctgc tgggcaaggt ggtggccctg cccgatcacc tgggcaaccg cctgcagcag
     2161 gagaacttgg ccgagttctt cccccagaac tacttccgcc tgctcggcga ggaggtcgtc
     2221 cgggtgctgc aggcggttgt ggactctctc caaggtggcc tggattcctc cgtgtccttc
     2281 gtgtctcagg tccttgggaa agcctgtgtc cacgggaggc agcaggagat cctgggcgtg
     2341 ctggtacccc ggctggcagc gctcacccag ggcagctacc tgcaccagcg cgtctgctgg
     2401 cgcctggtgg agcaagtgcc ggaccgggcc atggaggctg tgctgaccgg gctggtggag
     2461 gccgcactgg ggcctgaggt cctttcgaga ctgctgggga acctggtggt gaagaacaag
     2521 aaggcccagt ttgtgatgac ccagaagctt ctgttcttac agtcccggct cacgacgccc
     2581 atgctgcaga gcctgctggg ccatctggcc atggacagcc agcggcgccc gctcctgctg
     2641 caggtgctga aggagctgtt ggagacgtgg ggcagcagca gtgccatccg ccacactccc
     2701 ctgccgcagc agcgccacgt cagcaaggct gtcctcatct gcctggcgca actcggggag
     2761 ccggaactgc gggacagccg ggatgaactg ctggccagca tgatggcggg cgtgaagtgc
     2821 cgcctggaca gtagcctgcc ccccgtgcga cgcctgggca tgatcgtggc agaggtcgtt
     2881 agtgcccgga tccaccccga ggggcctccc ctgaaattcc agtacgaaga ggatgaactg
     2941 agcctcgagc tgctggcctt ggcctccccc cagcctgcgg gtgacggcgc ctcggaggcg
     3001 ggcacgtccc tcgttccagc cacggcagag ccccctgcag agacccccgc agagatcgtg
     3061 gatggcggcg tcccccaagc acagctggcg ggctctgact cggacctgga cagcgatgat
     3121 gagtttgtcc cctacgacat gtcgggggac agagagctga agagcagcaa ggctcctgcc
     3181 tacgtccggg actgcgtgga agccctgacc acgtctgagg acatagagcg ctgggaggca
     3241 gccctgcggg cccttgaggg cctggtctac aggagcccca cagccactcg ggaggtgagc
     3301 gtggagctgg ccaaggtgct tctgcatctg gaggagaaga cctgtgtggt gggatttgca
     3361 gggctgcgcc agagagccct ggtggccgtc acggtcacag acccggcccc ggtggccgac
     3421 tatctgacct cacagttcta tgccctcaac tacagcctcc ggcagcgcat ggacatcctg
     3481 gatgtgctga ctctggctgc ccaggagctg tctaggcctg ggtgcctcgg gaggactccc
     3541 caacctggct ccccaagtcc caacaccccg tgcctgccag aggcagccgt ctctcagcct
     3601 ggcagtgccg tggcgtctga ctggcgggtg gtggtggagg agcggatcag aagcaagacc
     3661 cagcggctct ccaagggtgg cccgaggcag ggcccggcag gcagccccag cagattcaac
     3721 tccgtggccg gccacttctt cttccccctc cttcagcgct ttgacaggcc tctggtgacc
     3781 ttcgacctct tgggagaaga ccagctggtt ctcggaaggc tggcgcacac cttaggggcc
     3841 ctgatgtgcc tggctgttaa caccacggtg gctgtggcca tgggcaaggc cctgctggaa
     3901 ttcgtgtggg cccttcgctt ccacatcgat gcctacgtgc gccaggggct gttgtcggcc
     3961 gtctcctccg tcctgctcag cctgcctgct gcgcgcctgc tggaggacct gatggacgag
     4021 ctgctggaag cccggtcctg gctggcggac gtggctgaga aagacccgga cgaggactgc
     4081 aggacgctgg cactgagggc cctgctgctt ctgcagagac tcaagaacag gctcctccca
     4141 cccgcgtctc cctagggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg
     4201 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4261 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4321 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4381 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4441 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4921 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     5041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     5101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5221 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5281 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5341 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5401 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5461 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5521 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5581 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5641 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5701 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5761 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5821 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5881 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5941 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6001 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6061 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6121 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6181 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6301 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6361 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6421 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6481 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6541 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6601 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6661 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6721 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6781 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6841 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7381 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     7441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     8641 agatctcccg atcccctatg gt
//
