LOCUS       V024933                 8215 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024933
VERSION     V024933
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..3705
                     /label="USP49(NM_001286554)"
                     /note="USP49(NM_001286554)"
                     /gene="USP49"
     polyA_signal    3782..4006
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4052..4480
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4494..4823
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4890..5684
                     /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    5858..5979
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6028..6044)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6052..6068
                     /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(6076..6106)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6121..6142
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6430..7015)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7186..8046)
                     /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(8047..8151)
                     /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 catggataga tgcaaacatg tagggcggtt acggctcgcc
     1681 caggaccact ccatcctgaa ccctcagaag tggtgctgct tagagtgtgc caccaccgag
     1741 tccgtgtggg cctgcctcaa gtgctcccac gtggcctgcg gccgctatat tgaggaccac
     1801 gccctgaaac actttgagga gacgggacac ccgctagcca tggaagtccg ggatctctac
     1861 gtgttctgtt acctgtgcaa ggactacgtg ctcaatgata acccagaggg ggacctgaag
     1921 ctgctaagaa gctccctcct ggcggtccgg ggccagaaac aggacacgcc ggtgagacgt
     1981 gggcggacgc tgcggtccat ggcttcgggt gaggacgtgg tcctgccgca gcgcgctcct
     2041 cagggacagc cgcagatgct cacggctctg tggtaccggc gtcagcgcct gctggccagg
     2101 acgctgcggc tgtggttcga gaagagctcc cggggccagg cgaagctgga gcagcggcgg
     2161 caggaggagg ccctggagcg caagaaggag gaggcgcgga ggcggcggcg cgaggtgaaa
     2221 cggcggctgc tggaggagct ggccagcacc cctccgcgca agagtgcacg gctgctcctg
     2281 cacacgcccc gcgacgcggg cccggctgcc tcgcgccccg ccgccctccc tacctcacgc
     2341 agagtgcccg ccgccacact caagctgcgt cgccagccgg ccatggcccc aggcgtcacg
     2401 ggcctgcgca acctgggcaa cacctgctac atgaactcca tcctccaggt gctcagccac
     2461 ctccagaagt tccgagaatg tttcctcaac cttgaccctt ccaaaacgga acatctgttt
     2521 cccaaagcca ccaacgggaa gactcagctt tctggcaagc caaccaacag ctcggccacg
     2581 gagctgtcct tgagaaatga cagggccgag gcatgcgagc gggagggctt ctgctggaac
     2641 ggcagggcct ccattagtcg gagtctggag ctcatccaga acaaggagcc gagttcaaag
     2701 cacatttccc tctgccgtga actgcacacc ctcttccgag tcatgtggtc cgggaagtgg
     2761 gccctagtgt cgcccttcgc catgctccac tcagtgtgga gcctgatccc tgccttccgc
     2821 ggctacgacc aacaggacgc gcaggaattt ctctgcgagc tgctgcacaa ggtgcagcag
     2881 gaactcgagt ctgagggcac cacacgccgg atcctcatcc ccttctccca gaggaagctc
     2941 accaaacagg tcttaaaggt ggtgaatacc atatttcatg ggcagctgct cagtcaggtc
     3001 acatgtatat catgcaatta caaatccaat accattgagc ccttttggga cctatccctg
     3061 gaattccctg aacgctatca ctgcatagaa aaggggtttg tccctttgaa tcaaacagag
     3121 tgcttgctca ctgagatgct ggccaaattc acagagacag aggccctgga agggagaatc
     3181 tacgcttgtg accagtgtaa cagcaaacga cgaaaatcca atcccaaacc ccttgttctg
     3241 agtgaagcta gaaagcagtt aatgatctac agactacctc aggttctccg gctgcacctt
     3301 aaaagattca ggtggtctgg ccgtaatcat cgagagaaga ttggggtcca tgtcgtcttt
     3361 gaccaggtat taaccatgga accttactgc tgcagggaca tgctctcctc tcttgacaaa
     3421 gagacctttg cctatgatct ctccgcagtg gtcatgcatc acgggaaagg gtttggctca
     3481 ggacactaca cagcctattg ctacaacaca gagggaggtt tttgggtcca ctgcaatgac
     3541 tcaaagctga atgtatgcag tgtcgaggaa gtgtgcaaaa cccaggccta catccttttt
     3601 tacactcaaa gaacagtgca gggcaatgca agaatctcag aaacccatct ccaagctcag
     3661 gtgcagtcca gcaacaatga tgaaggcaga ccacagacat tttcctgagg atcctgatat
     3721 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3781 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3841 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3901 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3961 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4021 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4381 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4501 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4621 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4681 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4741 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4801 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4861 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5041 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5401 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5701 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5761 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5821 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5881 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5941 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6001 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6061 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6121 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6241 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6961 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8161 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
