LOCUS       V019866                 7352 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019866
VERSION     V019866
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             902..1618
                     /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             1646..2878
                     /label="PACC1(NM_001198862)"
                     /note="PACC1(NM_001198862)"
                     /gene="PACC1"
     polyA_signal    2919..3143
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3189..3617
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3631..3960
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4027..4821
                     /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    4995..5116
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5165..5181)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5189..5205
                     /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(5213..5243)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5258..5279
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5567..6152)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6323..7183)
                     /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(7184..7288)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      961 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1021 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1081 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1141 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1201 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1261 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1321 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1381 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1441 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1501 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1561 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagct
     1621 cgagggagga gggggaagcg ccaccatgat ccggcaggag cgctccacat cctaccagga
     1681 ggctgtgcgt ccagcgcttc cttcaagcaa gccctgcctc ctcacttctc cagctgtatt
     1741 agtgaaactg ctctcctcct ccgcctccac ttctcggccc ccaaaccttg gtcatctttg
     1801 gcaaccgtcc tcttctgtgc ccctccatcg tgccgcctca ttggccaaag tcaggcagtt
     1861 ccagctgagt gaggagttgg tccaggtggt tgagaactca gagctggcag acgagcagga
     1921 caaggagacg gtcagagtcc aaggtccggg tatcttacca ggcctggaca gcgagtccgc
     1981 ctccagcagc atccgcttca gcaaggcctg cctgaagaac gtcttctcgg tcctactcat
     2041 cttcatctac ctgctgctca tggctgtggc cgtcttcctg gtctaccgga ccatcacaga
     2101 ctttcgtgag aaactcaagc accctgtcat gtctgtgtct tacaaggaag tggatcgcta
     2161 tgatgcccca ggtattgcct tgtaccccgg tcaggcccag ttgctcagct gtaagcacca
     2221 ttacgaggtc attcctcctc tgacaagccc tggccagccg ggtgacatga attgcaccac
     2281 ccagaggatc aactacacgg accccttctc caatcagact gtgaaatctg ccctgattgt
     2341 ccaggggccc cgggaagtga aaaagcggga gctggtcttc ctccagttcc gcctgaacaa
     2401 gagtagtgag gacttcagcg ccattgatta cctcctcttc tcttctttcc aggagttcct
     2461 gcaaagccca aacagggtag gcttcatgca ggcctgtgag agtgcctatt ccagctggaa
     2521 gttctctggg ggcttccgca cctgggtcaa gatgtcactg gtaaagacca aggaggagga
     2581 tgggcgggaa gcagtggagt tccggcagga gacaagtgtg gttaactaca ttgaccagag
     2641 gccagctgcc aaaaaaagtg ctcaattgtt ttttgtggtc tttgaatgga aagatccttt
     2701 catccagaaa gtccaagata tagtcactgc caatccttgg aacacaattg ctcttctctg
     2761 tggcgccttc ttggcattat ttaaagcagc agagtttgcc aaactgagta taaaatggat
     2821 gatcaaaatt agaaagagat accttaaaag aagaggtcag gcaacgagcc acataagcta
     2881 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2941 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3001 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3061 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3121 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3181 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3421 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3481 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3541 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3601 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3961 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4801 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4861 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4921 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4981 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5101 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5161 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5221 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5281 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5341 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5401 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5461 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5521 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5581 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5641 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5701 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5761 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5821 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5881 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5941 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6001 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6061 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6121 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6181 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6241 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6301 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6361 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6421 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6481 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6541 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6601 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6661 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6721 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6781 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6841 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6901 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6961 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7021 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7081 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7141 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7201 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7261 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7321 acggatcggg agatctcccg atcccctatg gt
//
