LOCUS       V033369                 9523 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033369
VERSION     V033369
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..5013
                     /label="USP36(NM_001385169)"
                     /note="USP36(NM_001385169)"
                     /gene="USP36"
     polyA_signal    5090..5314
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5360..5788
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5802..6131
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6198..6992
                     /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    7166..7287
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7336..7352)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7360..7376
                     /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(7384..7414)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7429..7450
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7738..8323)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8494..9354)
                     /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(9355..9459)
                     /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 catgccaata gtggataagt tgaaggaggc cctgaaaccc
     1681 ggccgcaagg actcggctga tgatggagaa ctggggaagc ttcttgcctc ctctgccaag
     1741 aaggtccttt tacagaaaat cgagttcgag ccagccagca agagcttctc ctaccagctg
     1801 gaggccttaa agagcaaata tgtgttgctc aaccccaaaa cagagggagc tagtcgccac
     1861 aagagtggag atgacccacc ggccaggaga cagggcagtg agcacacgta tgagagctgt
     1921 ggtgacggag tcccagcccc gcagaaagtg cttttcccca cggagcgact gtctctgagg
     1981 tgggagcggg tcttccgcgt gggcgcagga ctccacaacc ttggcaacac ctgctttctc
     2041 aatgccacca tccagtgctt gacctacaca ccacctctag ccaactacct gctctccaag
     2101 gagcatgctc gcagctgcca ccagggaagc ttctgcatgc tgtgtgtcat gcagaaccac
     2161 attgtccagg ccttcgccaa cagcggcaac gccatcaagc ccgtctcctt catccgagac
     2221 ctgaaaaaga tcgcccgaca cttccgcttt gggaaccagg aggacgcgca tgagttcctg
     2281 cggtacacca tcgacgccat gcagaaagcc tgcctgaatg gctgtgccaa gttggatcgt
     2341 caaacgcagg ctactacctt ggtccatcaa atttttggag ggtatctcag atcacgcgtg
     2401 aagtgctccg tgtgcaagag cgtctcggac acctacgacc cctacttgga cgtcgcgctg
     2461 gagatccggc aagctgcgaa tattgtgcgt gctctggaac tttttgtgaa agcagatgtc
     2521 ctgagtggag agaatgccta catgtgtgct aaatgcaaga agaaggttcc agccagcaag
     2581 cgcttcacca tccacagaac atccaacgtc ttaacccttt ccctcaagcg ctttgccaac
     2641 ttcagcgggg ggaagatcac caaggatgta ggctatccgg aattcctcaa catacgtccg
     2701 tatatgtccc agaataatgg tgatcctgtc atgtatggac tctatgctgt cctggtgcac
     2761 tcgggctaca gctgccatgc cgggcactat tactgctacg tgaaggcaag caatggacag
     2821 tggtaccaga tgaatgattc cttggtccat tccagcaacg tcaaggtggt tctgaaccag
     2881 caggcctacg tgctgttcta tctgcgaatt ccaggctcta agaaaagtcc cgagggcctc
     2941 atctccagga caggctcctc ctcccttccc ggccgcccga gtgtgattcc agatcactcc
     3001 aagaagaaca tcggcaatgg gattatttcc tccccactga ctggaaagcg acaagactct
     3061 gggacgatga agaagccgca caccactgaa gagattggtg tgcccatatc caggaatggc
     3121 tccaccctgg gcctgaagtc ccagaacggc tgcattcctc caaagctgcc ctcggggtcc
     3181 ccttccccca aactctccca gacacccaca cacatgccaa ccatcctaga cgaccctgga
     3241 aagaaggtga agaagccagc tcctccacag cacttttccc ccagaactgc tcaggggctg
     3301 cctgggacca gcaactcgaa tagcagcaga tctgggagcc aaaggcaggg ctcctgggac
     3361 agcagggatg ttgtcctctc tacctcacct aagctcctgg ctacagccac tgccaacggg
     3421 catgggctga aggggaacga cgagagcgct ggcctcgaca ggaggggctc cagcagctcc
     3481 agcccagagc actcggccag cagcgactcc accaaggccc cccagacccc caggagtgga
     3541 gcggcccatc tctgcgattc tcaggaaacg aactgttcca ccgctggcca ctccaaaacg
     3601 ccgccaagtg gagcagattc taagacggtg aagctgaagt cccctgtcct gagcaacacc
     3661 accactgagc ctgcaagcac catgtctcct ccaccagcca aaaaactggc cctttctgcc
     3721 aagaaggcca gcaccctgtg gagggcgacc ggcaatgacc tccgtccacc tcccccctca
     3781 ccatcctccg acctcaccca ccccatgaaa acctctcacc ccgtcgttgc ctccacttgg
     3841 cccgtccata gagccagggc tgtgtcacct gctccccaat catccagccg cctgcaaccc
     3901 cccttcagcc cccaccccac attgctgtcc agtaccccca agcccccagg gacgtcagaa
     3961 ccacggagct gctcctccat ctcgacggcg ctgcctcagg tcaacgagga ccttgtgtct
     4021 cttccacacc agttgccaga ggccagtgag cccccccaga gcccctctga gaagaggaaa
     4081 aagacctttg tgggagagcc gcagaggctg ggctcagaga cgcgcctccc acagcacatc
     4141 agggaggcca ctgcggctcc ccacgggaag aggaagagga agaagaagaa gcgcccggag
     4201 gacacagctg ccagcgccct gcaggagggg cagacacaga gacagcctgg gagccccatg
     4261 tacaggaggg agggccaggc acagctgccc gctgtcagac ggcaggaaga tggcacacag
     4321 ccacaggtga atggccagca ggtgggatgt gttacggacg gccaccacgc gagcagcagg
     4381 aagcggagga ggaaaggagc agaaggtctt ggtgaagaag gcggcctgca ccaggaccca
     4441 cttcggcaca gcagctgctc tcccatgggt gatggtgatc cagaggccat ggaagagtct
     4501 ccaaggaaaa agaaaaagaa aaaaagaaag caggagacac agcgggcagt agaagaggat
     4561 gggcatctca aatgcccaag gagtgccaag ccccaagatg ctgttgtccc cgagtccagc
     4621 agctgcgcac catccgcgaa tggctggtgt cctggggacc gcatggggct gagccaggcc
     4681 cctcctgtgt cttggaatgg agagcgggag tctgatgtgg tccaggaact gctcaaatac
     4741 tcatctgata aagcttacgg gagaaaagtt ctgacctggg atggcaagat gtcggcggtc
     4801 agtcaggatg ctattgaaga cagcagacag gcccggactg agaccgtggt tgatgactgg
     4861 gacgaagagt ttgaccgagg gaaggaaaag aaaattaaaa aatttaagag agagaagagg
     4921 agaaacttca acgccttcca gaaacttcag actcgacgga acttctggtc tgtgactcac
     4981 ccagcaaagg ctgccagcct cagctatcgc cgctgaggat cctgatatcc agcacagtgg
     5041 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     5101 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5161 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5221 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     5281 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     5341 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5461 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5521 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5581 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5641 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5701 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5761 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     5821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6061 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6121 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     6181 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     6241 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     6301 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     6361 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     6421 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     6481 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     6541 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     6601 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     6661 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     6721 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     6781 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     6841 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     6901 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     6961 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     7021 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7081 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7141 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7201 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7261 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     8281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9481 acctgacgtc gacggatcgg gagatctccc gatcccctat ggt
//
