LOCUS       V042754                 7483 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042754
VERSION     V042754
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..2973
                     /label="Tubb5(NM_173102)"
                     /note="Tubb5(NM_173102)"
                     /gene="Tubb5"
     polyA_signal    3050..3274
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3320..3748
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3762..4091
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4158..4952
                     /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    5126..5247
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5296..5312)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5320..5336
                     /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(5344..5374)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5389..5410
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5698..6283)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6454..7314)
                     /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(7315..7419)
                     /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 catgagggaa atcgtgcaca tccaggccgg acagtgtggc
     1681 aaccagatcg gtgctaagtt ctgggaggtg ataagcgatg aacacggcat cgaccccacc
     1741 ggcacctacc acggagacag cgacttgcag ctggaccgaa tctctgtgta ctacaatgaa
     1801 gctacaggtg gcaagtatgt ccctcgagct atcttagtgg atctagaacc cgggactatg
     1861 gactccgttc gctcaggtcc ttttggccag atcttcagac cggacaactt tgtttttggt
     1921 cagtctgggg caggcaacaa ctgggctaag ggtcactaca cagagggagc tgagctggtt
     1981 gactctgtct tggatgtggt gcggaaggag gcggagagct gtgactgcct gcaaggcttt
     2041 cagctgaccc attcgctggg tggaggcacg ggctctggca tgggcaccct gctcatcagc
     2101 aagattcgag aagaataccc cgaccgcatc atgaacacct ttagcgtggt gccctcaccc
     2161 aaagtgtctg acaccgtggt tgagccctac aacgccaccc tgtccgtcca ccagttggta
     2221 gagaacacag atgagaccta ctgcattgac aacgaggccc tctacgacat ctgcttccgt
     2281 acccttaagc tcaccacgcc aacctacgga gacctgaacc atctcgtctc tgccaccatg
     2341 agcggtgtca ccacctgcct ccgtttcccg ggccagctga acgccgacct tcggaagctg
     2401 gctgtcaaca tggtgccctt cccgcgtctc cacttcttta tgcctggctt tgcccctctc
     2461 accagccgtg gaagccagca gtaccgggcc ctcactgtgc ctgaactcac ccagcaggtc
     2521 ttcgacgcca agaacatgat ggccgcctgt gacccacgcc acggccggta cctcacggtt
     2581 gcggctgtct tccgtggacg gatgtccatg aaggaggtag atgagcaaat gctcaacgtg
     2641 cagaataaga atagcagcta cttcgtggaa tggatcccca acaacgtcaa gacggctgtc
     2701 tgtgacatcc caccgcgagg cctcaagatg gcagtcacct tcatcggaaa cagcacggcc
     2761 atccaggagc tcttcaagcg catctctgag cagtttacgg caatgttccg tcggaaggct
     2821 ttcctccact ggtacacggg tgagggcatg gacgagatgg agttcaccga agctgagagc
     2881 aacatgaatg acctcgtctc tgagtatcag cagtaccagg acgccaccgc agaagaggaa
     2941 gaggatttcg gagaggaggc agaagaagag gcctaaggat cctgatatcc agcacagtgg
     3001 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3061 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3121 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3181 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3241 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3301 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3361 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3421 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3481 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3541 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3601 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3661 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3721 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3781 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3841 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3901 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3961 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4021 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4081 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     4141 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4201 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4261 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4321 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4381 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4441 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4501 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4561 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4621 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4681 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4741 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4801 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4861 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4921 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4981 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5041 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5101 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5161 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5221 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5281 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5341 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5401 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7441 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
