LOCUS       V020197                 8204 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020197
VERSION     V020197
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..3730
                     /label="APP(NM_201414)"
                     /note="APP(NM_201414)"
                     /gene="APP"
     polyA_signal    3771..3995
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4041..4469
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4483..4812
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4879..5673
                     /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    5847..5968
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6017..6033)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6041..6057
                     /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(6065..6095)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6110..6131
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6419..7004)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7175..8035)
                     /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(8036..8140)
                     /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 ccaccatgct gcccggtttg gcactgctcc tgctggccgc
     1681 ctggacggct cgggcgctgg aggtacccac tgatggtaat gctggcctgc tggctgaacc
     1741 ccagattgcc atgttctgtg gcagactgaa catgcacatg aatgtccaga atgggaagtg
     1801 ggattcagat ccatcaggga ccaaaacctg cattgatacc aaggaaggca tcctgcagta
     1861 ttgccaagaa gtctaccctg aactgcagat caccaatgtg gtagaagcca accaaccagt
     1921 gaccatccag aactggtgca agcggggccg caagcagtgc aagacccatc cccactttgt
     1981 gattccctac cgctgcttag ttggtgagtt tgtaagtgat gcccttctcg ttcctgacaa
     2041 gtgcaaattc ttacaccagg agaggatgga tgtttgcgaa actcatcttc actggcacac
     2101 cgtcgccaaa gagacatgca gtgagaagag taccaacttg catgactacg gcatgttgct
     2161 gccctgcgga attgacaagt tccgaggggt agagtttgtg tgttgcccac tggctgaaga
     2221 aagtgacaat gtggattctg ctgatgcgga ggaggatgac tcggatgtct ggtggggcgg
     2281 agcagacaca gactatgcag atgggagtga agacaaagta gtagaagtag cagaggagga
     2341 agaagtggct gaggtggaag aagaagaagc cgatgatgac gaggacgatg aggatggtga
     2401 tgaggtagag gaagaggctg aggaacccta cgaagaagcc acagagagaa ccaccagcat
     2461 tgccaccacc accaccacca ccacagagtc tgtggaagag gtggttcgag ttcctacaac
     2521 agcagccagt acccctgatg ccgttgacaa gtatctcgag acacctgggg atgagaatga
     2581 acatgcccat ttccagaaag ccaaagagag gcttgaggcc aagcaccgag agagaatgtc
     2641 ccaggtcatg agagaatggg aagaggcaga acgtcaagca aagaacttgc ctaaagctga
     2701 taagaaggca gttatccagc atttccagga gaaagtggaa tctttggaac aggaagcagc
     2761 caacgagaga cagcagctgg tggagacaca catggccaga gtggaagcca tgctcaatga
     2821 ccgccgccgc ctggccctgg agaactacat caccgctctg caggctgttc ctcctcggcc
     2881 tcgtcacgtg ttcaatatgc taaagaagta tgtccgcgca gaacagaagg acagacagca
     2941 caccctaaag catttcgagc atgtgcgcat ggtggatccc aagaaagccg ctcagatccg
     3001 gtcccaggtt atgacacacc tccgtgtgat ttatgagcgc atgaatcagt ctctctccct
     3061 gctctacaac gtgcctgcag tggccgagga gattcaggat gaagttgatg agctgcttca
     3121 gaaagagcaa aactattcag atgacgtctt ggccaacatg attagtgaac caaggatcag
     3181 ttacggaaac gatgctctca tgccatcttt gaccgaaacg aaaaccaccg tggagctcct
     3241 tcccgtgaat ggagagttca gcctggacga tctccagccg tggcattctt ttggggctga
     3301 ctctgtgcca gccaacacag aaaacgaagt tgagcctgtt gatgcccgcc ctgctgccga
     3361 ccgaggactg accactcgac caggttctgg gttgacaaat atcaagacgg aggagatctc
     3421 tgaagtgaat ctggatgcag aattccgaca tgactcagga tatgaagttc atcatcaaaa
     3481 attggtgttc tttgcagaag atgtgggttc aaacaaaggt gcaatcattg gactcatggt
     3541 gggcggtgtt gtcatagcga cagtgatcgt catcaccttg gtgatgctga agaagaaaca
     3601 gtacacatcc attcatcatg gtgtggtgga ggttgacgcc gctgtcaccc cagaggagcg
     3661 ccacctgtcc aagatgcagc agaacggcta cgaaaatcca acctacaagt tctttgagca
     3721 gatgcagaac tagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3781 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3841 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3901 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3961 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4021 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4081 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4141 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4201 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4261 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4321 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4381 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4441 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     4501 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4561 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4621 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     4681 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4741 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4801 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4861 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4921 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4981 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5041 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     5101 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5161 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5221 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5281 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5341 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5401 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5461 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5521 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5581 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5641 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5701 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5761 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5821 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5881 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5941 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6001 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6061 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6121 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6181 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6241 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6301 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6361 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6421 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6481 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6541 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6601 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6661 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6721 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6781 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6841 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6901 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     7021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     7081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     7141 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7201 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     7261 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     7321 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     7381 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     7441 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     7501 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     7561 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     7621 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     7681 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     7741 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7801 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7861 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7921 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7981 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8041 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     8101 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     8161 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggt
//
