LOCUS       V033071                 8557 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033071
VERSION     V033071
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..4047
                     /label="CDC5L(NM_001253)"
                     /note="CDC5L(NM_001253)"
                     /gene="CDC5L"
     polyA_signal    4124..4348
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4394..4822
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4836..5165
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5232..6026
                     /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    6200..6321
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6370..6386)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6394..6410
                     /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(6418..6448)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6463..6484
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6772..7357)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7528..8388)
                     /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(8389..8493)
                     /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 catgcctcga attatgatca aggggggcgt atggaggaat
     1681 accgaggatg aaattctgaa agcagcggta atgaaatatg ggaaaaatca gtggtctagg
     1741 attgcctcat tgctgcatag aaaatcagca aagcagtgca aagccagatg gtatgaatgg
     1801 ctggatccaa gcattaagaa gacagaatgg tccagagaag aagaggaaaa actcttgcac
     1861 ttggccaagt tgatgccaac tcagtggagg accattgctc caatcattgg aagaacagcg
     1921 gcccagtgct tagaacacta tgaatttctt ctggataaag ctgcccaaag agacaatgaa
     1981 gaggaaacaa cagatgatcc acgaaaactt aaacctggag aaatagatcc aaatccagaa
     2041 acaaaaccag cgcggcctga tccaattgat atggatgagg atgaacttga gatgctttct
     2101 gaagccagag cccgcttggc taatactcag ggaaagaagg ccaagaggaa agcaagagag
     2161 aaacaattgg aagaagcaag acgtcttgct gccctccaaa aaagaagaga acttcgagca
     2221 gctggcatag aaattcagaa gaaaagaaaa aggaagagag gagttgatta taatgccgaa
     2281 atcccatttg aaaaaaagcc tgcccttggt ttttatgata cttctgagga aaactaccaa
     2341 gctcttgacg cagatttcag gaaattaaga caacaggatc ttgatgggga gctaagatct
     2401 gaaaaagaag gaagagatag aaaaaaagac aaacagcatt tgaaaaggaa aaaagaatct
     2461 gatttaccat cagctattct tcaaactagt ggtgtttctg aatttactaa aaagagaagc
     2521 aaactagtac ttcctgcccc tcagatttca gatgcagaac tccaggaagt tgtaaaagta
     2581 ggccaagcga gtgaaattgc acgtcaaact gccgaggaat ctggcataac aaattctgct
     2641 tccagtacac ttttgtctga gtacaatgtc accaacaaca gcgttgctct tagaacacca
     2701 cgaacaccag cttcccagga cagaattctg caggaagccc agaacctcat ggccctcacc
     2761 aatgtggaca ccccattgaa aggtggactt aataccccat tgcatgagag tgacttctca
     2821 ggtgtaactc cacagcgaca agttgtacag actccaaaca cagttctctc tactccattc
     2881 aggactcctt ctaatggagc tgaagggctg actccccgga gtggaacaac tcccaaacca
     2941 gttattaact ctactccggg tagaactcct cttcgagaca agttaaacat taatcccgag
     3001 gatggaatgg cagactatag tgatccctct tacgtgaagc agatggaaag agaatcccga
     3061 gaacatctcc gtttagggtt gttgggcctt cctgccccta agaatgattt tgaaattgtt
     3121 ctaccagaaa atgccgagaa ggagctggaa gaacgtgaaa tagatgatac ttacattgaa
     3181 gatgctgctg atgtggatgc tcgaaagcag gccatacgag atgcagagcg tgtaaaggaa
     3241 atgaaacgaa tgcataaagc tgtccagaaa gatctgccaa gaccatcaga agtaaatgaa
     3301 actattctaa gacccttaaa tgtagaaccg cctttaacag atttacagaa aagtgaagaa
     3361 ctaatcaaaa aagaaatgat cacaatgctt cattatgacc ttctacatca cccttatgaa
     3421 ccatctggaa ataaaaaagg caaaactgta gggtttggta ccaataattc agagcacatt
     3481 acctatctgg aacataatcc ttatgaaaag ttctccaaag aagagctgaa aaaggcccag
     3541 gatgttttgg tgcaggagat ggaagtggtt aaacaaggaa tgagccatgg agagctctca
     3601 agtgaagctt ataaccaggt gtgggaagaa tgctacagtc aagttttata tcttcctggg
     3661 cagagccgct acacacgggc caatctggct agtaaaaagg acagaattga atcacttgaa
     3721 aagaggctcg agataaacag gggtcacatg acgacagaag ccaagagggc tgcaaagatg
     3781 gaaaagaaga tgaaaatttt gcttgggggt taccagtctc gtgctatggg gctcatgaaa
     3841 cagttgaatg acttatggga ccaaattgaa caggctcact tggagttacg cacttttgaa
     3901 gaactcaaga aacatgaaga ttctgctatt ccccggaggc tagagtgtct aaaagaagac
     3961 gttcagcgac aacaagaaag agaaaaggaa cttcaacata gatatgctga tttgctgctg
     4021 gagaaagaga ctttaaagtc aaaattctga ggatcctgat atccagcaca gtggcggccg
     4081 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     4141 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4201 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4261 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4321 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4381 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4441 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4501 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4561 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4621 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4681 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4741 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4801 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4861 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4921 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4981 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5041 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5101 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5161 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5221 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5281 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5341 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5401 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5461 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5521 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5581 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5641 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5701 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5761 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5821 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5881 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5941 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6001 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6061 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6121 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6181 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6241 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6301 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6361 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6421 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6481 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6541 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6601 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6661 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6721 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6781 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6841 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6901 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6961 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     7021 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     7081 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7141 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7201 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     7261 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     7321 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7381 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7441 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7501 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7561 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7621 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7681 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7741 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7801 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7861 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7921 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7981 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8041 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8101 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8161 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8221 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8281 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8341 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8401 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8461 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8521 cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
