LOCUS       V044183                 9442 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044183
VERSION     V044183
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..4101
                     /label="Usp43(NM_173754)"
                     /note="Usp43(NM_173754)"
                     /gene="Usp43"
     CDS             4108..4134
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4138..4164
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4168..4194
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    4218..4806
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4872..4920
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5122..5550
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5564..5893
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5960..6754
                     /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    6930..7051
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7100..7116)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7124..7140
                     /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(7148..7178)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7193..7214
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7502..8090)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8261..9121)
                     /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(9122..9226)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgga tccgggcgtg
      721 gggaacgcac tgggagaggg gccgcccgcg ccccggcccc gccgccgccg ttcccttcgc
      781 cgcctgctta accgcttcct gctagctctg ggcagccgct cccgctcggg ggactcgccg
      841 ccccggcccc cggcccagcc cagtccctac gatggcgatg gagaaggggg ctttgcctgc
      901 gccccgggcc cggcaccagc ctctgccggg agtccagggc cagatcgtcc tccgggatcc
      961 caaccccaga tatcttcggg tgacggggcg cgtccgcccg gcgctcaggg cttgaagaac
     1021 catggcaaca cctgtttcat gaacgcggtg gtgcagtgtc tcagcaacac cgacctgctg
     1081 gccgagttcc tagcgctggg gcgctaccgg gcggcgccgg gccgcgccga ggtcaccgag
     1141 cagctggcgg cgctggtgcg cgcgctctgg acgcgcgaat acacgcccca actttccgca
     1201 gagttcaaga atgctgtttc taagtatggc tcccagttcc aaggcaattc ccagcacgat
     1261 gccctggaat tcctgctctg gcttctggat cgtgtccacg aggacttgga aggctcagct
     1321 cacgggctgg tatctgagca gctgccccca gaagtgagta agatctcaga ggacctccgg
     1381 ccatctgcag ctcctacctc cttagggccc agctttgtgc agagccactt tcaagcacag
     1441 tacaggtctt ccttgacttg tccacactgc ctaaaacaaa gcaacacctt cgaccctttc
     1501 ctgtgtgtgt cccttcccat ccccttgcgc cagacgagat tcttgagtgt cactttggtg
     1561 ttcccctcaa agagccagcg gtttctgcgt gttggcttgg ccgtgccaat cctcagcaca
     1621 gtggcggcct tgaggaagat ggttgcagag gaaggaggtg tccctgcaga ggaggtgatc
     1681 ttggttgaac tgtatcccaa tggattccag cggtcattct ttgatgaaga ggacctgaat
     1741 accattgcag agggtgacaa tgtgtatgct ttccaagtcc ctccttcacc tggtctgggg
     1801 acgctctcag cccatccgtc tggtctgtca gtgtcaccgc gcttgccagt ccgtgacagc
     1861 cagcgattct ctgggcctct ccacagcgag aacagggtgg tctttctctt ctgtaacctg
     1921 gtgggctcag ggcagcaggc gagcaggttt ggaccaccgt tcttgattcg ggaagataga
     1981 acaatctcct gggcccagct tcaacagtgt attctcagca aggttcgatg cctcatgagg
     2041 agtgaggtct ctgcacagga cctggggact ctgttctcca tccgggttgt ggggctctcc
     2101 ttggcctgca gctacttatc tccaaaggac aaccggcccc tttgccactg ggcagttgac
     2161 agggctttgc acctcaggag gccaggaggc cctcctcatg tcaagctggc tgtggaatgg
     2221 gacagctctg tcacagagcg cctgtttggg agcctccagg aggaacgggt ccaggatgca
     2281 gacagtgtgt ggcggcagca gcaggcacac cagcagccca gctgtaccct ggacgaatgt
     2341 tttcagtcct ataccaagga ggaacagctg gcccaggatg atgcatggaa gtgccctcac
     2401 tgccaagtcc tccagcaggg tgtggtcaag ctgagtctgt ggactctgcc cgacatcctg
     2461 atcatccatc tgaaacggtt ctgtcaggtg ggagagagaa ggaacaagct ctccacgctg
     2521 gtgaagttcc cactgtctgg actcaacatg gctccccatg tagcacggag gagcaccaac
     2581 tccaaggcgg ggcccggccc ttggtcctcc tggaagcagc cgatctgcct ccccaccacc
     2641 tacccgctgg acttcctcta cgacttgtat gctgtctgca accaccatgg cagtctgcaa
     2701 ggtgggcatt acacagccta ttgccggaac tccctggacg gccagtggta cagctatgac
     2761 gacagcacag tggaggccct gcgagaggat gaggtcaaca ccaggggggc ttacatccta
     2821 ttctaccaga agcggaacag catccctccc tggtcagcca gcagctccat gagaggttcc
     2881 accagctcct ccttgtctga tcattggctc atgagactcg ggagcctcaa caacagcaca
     2941 aggggaagct tgctgtcctg gagctcggcc ccctgcccct ccatggcccg ggtacccgac
     3001 tctcctgtct tcaccaatgg tgtctgccac caggacaagg gaggagttga gaccagacct
     3061 ttggttcggg gtgttggtgg ccgaagtatc agcatgaagg catctcctgc ttcccgatcc
     3121 aggcatgggc ccttcaagac catgcccctg aggtggtcat ttggacacag ggagaaacgt
     3181 cccggggcat ccgttgaact ggtggaatac ctagagtcca gaagaagacc ccgttctacc
     3241 agccagtcca ttgtgccgct cttgactcgt gctgctggtg gggaagagac ttcagcgtca
     3301 ccgaggtcag atggcaccct tcctgctaaa agtgaagaca gcggtcgagc cattgggcaa
     3361 ggaacaactg gggtgcccct ttcctcatgc cacctcaacc accatccagc cctggggtct
     3421 ttagatgaca gtctgcacac tgccaggaca cgaacgggga atgtaagtca ggatatcagg
     3481 ctcccaaaaa agtttgacct gcccctcact gtgatgccct cagtggggga tgaaaagcca
     3541 gctcgccctg agggccagaa gatgacacct tggaagggaa gtagccaggt gggaagccag
     3601 agtagcccac cttccccctc cactggcttg cttagaaact ttaaggacag tggcccaggt
     3661 accttaccca agatgaagtc taaggctgcc atggaggaaa gggcccctga caaagacagg
     3721 ggacagggga cattcactct tttgaagtct gtgttttgga agaaagaaca caagaggact
     3781 gtcaggactg agagctcacc accagcaccc ccaatctccc tagggagtga ccggctgagc
     3841 cctgctgcca tgaatgagca ggccctcagg attcgagaga gcccagccaa gggcctgggc
     3901 aaccacatgg agagggacat cagatctgcg cccagctctc tccacctccc ccgaaaagcc
     3961 agcaggcccc ctcgagccag tacagctggc acctcacaaa ggaccattcc tggggaacag
     4021 atttcttatg gcaccctgca gagagtgaaa taccacaccc tttccttagg tcgaaagaaa
     4081 tccttacctg agtccagctt cctcgagtac ccatacgacg tacctgatta tgcaacttat
     4141 ccttatgatg tacctgatta tgctagctat ccttatgatg tcccagacta cgcatagtga
     4201 gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4261 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4321 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4381 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4441 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4501 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4561 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     4621 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4681 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4741 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4801 ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     4861 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc
     4921 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4981 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5041 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct
     5101 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5161 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5221 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     5281 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5341 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5401 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5461 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     5521 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     5581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     5761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5821 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5881 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5941 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6001 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6061 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6121 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6181 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     6241 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     6301 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6361 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6421 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6481 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6541 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6601 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6661 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6721 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga
     6781 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6841 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6901 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6961 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7021 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     7081 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7141 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7201 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7261 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7321 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7381 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7441 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7501 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7561 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7621 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7681 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7741 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7801 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7861 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7921 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7981 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8041 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8101 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8161 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8221 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8281 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8341 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8401 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8461 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8521 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8581 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8641 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8701 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8761 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8821 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8881 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8941 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9001 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9061 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9121 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9181 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9241 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag
     9301 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     9361 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     9421 ttgcatgaag aatctgctta gg
//
