LOCUS       V028816                10024 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028816
VERSION     V028816
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..5514
                     /label="Per1(NM_011065)"
                     /note="Per1(NM_011065)"
                     /gene="Per1"
     polyA_signal    5591..5815
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5861..6289
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6303..6632
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6699..7493
                     /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    7667..7788
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7837..7853)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7861..7877
                     /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(7885..7915)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7930..7951
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8239..8824)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8995..9855)
                     /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(9856..9960)
                     /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 catgagtggt cccctagaag gggccgatgg gggaggagac
     1681 cccaggcccg gagaaccttt ttgtcctgga ggagtcccat cccctggggc cccgcagcac
     1741 cggccttgtc caggccccag cctggctgat gacactgatg caaacagcaa tggctcaagt
     1801 ggcaatgagt ccaacggacc cgagtccagg ggcgcatctc agcggagttc tcatagttcc
     1861 tcttctggca atggcaagga ctcagctctg ctggagacca ctgagagcag caagagtaca
     1921 aactcacaga gcccatcccc acccagcagc tccattgcct acagcctcct gagtgcgagc
     1981 tcagagcagg acaacccatc taccagtggc tgcagcagtg aacagtcagc tcgagccagg
     2041 acccagaaag aactcatgac tgcacttcgg gagctcaaac ttcgactgcc accagagcgt
     2101 cggggcaagg gccgctctgg gaccttggcc acactgcagt acgctctggc ctgtgtcaag
     2161 caggttcagg ctaaccagga atattaccag cagtggagtc tggaggaggg tgagccttgt
     2221 gccatggaca tgtctactta caccctggag gaattggagc atatcacatc cgaatacaca
     2281 cttcgaaacc aggacacctt ctctgtggct gtgtccttcc tgacaggccg gattgtctat
     2341 atttcggagc aggcaggtgt cctgctgcgt tgcaaacggg atgtgtttcg gggtgcccgc
     2401 ttctcagagc tcctggctcc ccaggatgtg ggtgtcttct atggctctac tacaccatct
     2461 cgactgccca cctggggcac tggcacctct gcaggttcag gtctcaagga cttcacccag
     2521 gaaaagtctg tcttctgccg aatcagagga ggtcctgacc gggatccagg gcctcggtac
     2581 cagccattcc gcctaacccc atatgtgacc aagattcggg tctcagatgg agcccctgca
     2641 cagccgtgct gcctactcat tgccgagcgc atccactctg gttatgaagc tccccggatc
     2701 cctcctgaca agaggatctt caccacccga cacacaccaa gctgcctctt ccaggatgta
     2761 gatgaaaggg ctgccccact gctgggttac cttccccagg atctcctggg ggctccagta
     2821 cttctctttc tacatcctga ggaccgaccc ctcatgctgg ccattcataa gaagatactg
     2881 cagctggcag gccagccctt tgaccattcc cctattcgct tctgtgctcg gaacggggaa
     2941 tatgtcacca tggacaccag ctgggccggt tttgtgcacc cctggagccg caaggtggct
     3001 ttcgtgttgg gtcgccataa agtgcgcacg gcacccctga atgaggacgt cttcactccc
     3061 ccagccccca gcccagctcc gtccctggac tctgatatcc aggagctctc agagcagatc
     3121 catcgattgc tgctgcagcc tgtgcacagc tccagcccca cggggctctg tggagttggc
     3181 cctctgatgt cccctggtcc tctacacagc cctggctcct ccagtgatag caatgggggg
     3241 gacgctgagg ggcctgggcc tcctgctcca gtgactttcc agcagatctg taaggatgtg
     3301 catctggtaa agcaccaggg acaacagctc ttcattgaat ctcgggccaa gcccccaccc
     3361 cggccccgcc tccttgctac aggtacattc aaagccaaag tccttccctg ccagtcccca
     3421 aaccccgaac tggaggtggc cccagttcct gaccaagcct cgttagcctt ggcccctgag
     3481 gagccagaga ggaaagaaac ctctggctgt tcctaccagc agatcaactg cctggacagc
     3541 atcctcaggt atttggagag ctgcaacatt cccagtacaa ccaagcgtaa atgtgcctcc
     3601 tcctcctcct acactgcctc ttcagcctct gatgatgaca agcagagggc aggtccagtt
     3661 cctgtggggg ccaagaaaga tccgtcgtca gcaatgctgt ctggggaggg ggcaactcct
     3721 cggaaggagc cagtggtggg aggcaccctg agcccgctcg ccctggccaa taaggcagag
     3781 agcgtggtgt ccgtcaccag tcagtgtagc ttcagctcca ccatcgtcca tgtgggagac
     3841 aagaagcccc cggagtcgga catcatcatg atggaagacc tgcctggcct ggcccctggc
     3901 ccagccccca gtccggcccc cagccccaca gtagcccctg acccaacccc agatgcttat
     3961 cgcccagtgg gtctgaccaa ggccgtgctg tccctgcaca cacagaagga agagcaagcc
     4021 ttcctcaacc gcttcagaga tcttggcagg cttcgtggac ttgacacctc ttctgtggcc
     4081 ccctcagccc ctggctgcca ccatggcccc attccccctg gtcgccgaca ccactgccga
     4141 tctaaagcaa agcgttcccg ccaccaccac caccagaccc cccggcccga aactccctgc
     4201 tatgtctccc atccttcacc tgtgccctct tctggaccct ggccaccccc accagccacg
     4261 acccccttcc cagcaatggt ccagccctac ccactcccag tattctcccc tcgaggagga
     4321 ccccagcccc ttccccctgc ccctacatct gtgtcccctg ctaccttccc ttctccctta
     4381 gtgaccccaa tggtggcctt ggtgctccct aactatctat tccctacccc acctagttat
     4441 ccatatgggg tgtcccaggc ccctgttgag gggccaccca cgcctgcttc ccactcgccc
     4501 tctccatccc tgcccccacc acctctcagc cccccccacc gcccagactc cccactgttc
     4561 aactcgagat gcagctcccc actccagctc aatctgctgc agcttgagga gtccccccgc
     4621 acggaggggg gcgctgctgc aggaggccca ggaagcagtg ctgggcccct gcctcccagt
     4681 gaggagactg ctgagccaga ggccagattg gtggaggtta ctgagtcgtc caatcaggat
     4741 gcactttcag gctccagcga cctgctggag ctactgctcc aagaagactc tcgctcgggc
     4801 acaggctccg cagcctcagg ctccctgggc tctggcctgg gctctgggtc tggttcagga
     4861 tcccacgaag ggggaagcac ctcagccagc atcacccgca gcagtcagag cagccataca
     4921 agcaagtact ttggcagcat cgactcttcc gaggctgaag ctggggctgc tcgggccagg
     4981 actgagcctg gggaccaggt cattaagtgt gtgctccagg accccatctg gctgctcatg
     5041 gccaatgccg accagcgtgt catgatgaca taccaggtgc cgtccaggga tgcagcctct
     5101 gtgctgaagc aagaccggga gaggctccgg gccatgcaga aacagcagcc acggttctca
     5161 gaggaccaga ggcgggaact gggtgctgtg cactcctggg tccggaaggg ccagctgcct
     5221 cgggcccttg atgtgacggc gtgtgtggac tgtggcagca gcgttcaaga tcctggccac
     5281 tctgatgacc ccctcttctc agaactggat ggattggggc tggagcccat ggaagagggt
     5341 ggaggcgagg gtggtgggtg tggtgttggc ggtggtgggg gtgatggtgg tgaggaggcc
     5401 cagacccaaa ttggggctaa gggttcaagc tctcaggact ctgccatgga ggaagaagag
     5461 caaggtgggg gctcatccag cccagcttta cctgcagaag aaaacagcac cagctaggga
     5521 tcctgatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     5581 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5641 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     5701 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     5761 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     5821 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     5881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     6301 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6361 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6421 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6481 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6541 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     6601 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     6661 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     6721 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     6781 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     6841 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     6901 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     6961 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     7021 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     7081 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     7141 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     7201 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     7261 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     7321 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     7381 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     7441 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     7501 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7561 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7621 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7681 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7741 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7801 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7921 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     8041 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8101 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8161 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8221 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8281 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8341 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8401 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8461 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8521 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8581 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8641 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8701 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8761 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8821 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8881 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8941 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9001 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9061 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9121 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9181 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9241 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9301 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9361 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9421 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9481 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9541 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9601 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9661 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9721 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9781 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9841 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9901 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9961 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    10021 tggt
//
