LOCUS       V035209                 8935 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035209
VERSION     V035209
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..3591
                     /label="Usp4(NM_011678)"
                     /note="Usp4(NM_011678)"
                     /gene="Usp4"
     CDS             3598..3687
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3711..4299
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4365..4413
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4615..5043
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5057..5386
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5453..6247
                     /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    6423..6544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6593..6609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6617..6633
                     /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(6641..6671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6686..6707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6995..7583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7754..8614)
                     /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(8615..8719)
                     /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 ccaccatggc ggaaggccgg
      721 ggcagccgtg agcgaccgga tgtggagacc cagaagacgg agctcggagc cttgatgggg
      781 accacgctcc aacgtggggc gcagtggtat cttattgaca gccggtggtt caagcagtgg
      841 aagaagtatg tgggctttga cagctgggac atgtacaatg tcggggagca taacctgttt
      901 cctggaccta ttgacaactc cggactcttc tcagatcctg agagtcagac cttgaaggag
      961 cacttaatcg atgagctgga ctatgtgctg gtcccagccg aagcctggaa taaattgctg
     1021 aattggtatg gctgtgtgga gggccagcag cctattgtca gaaaagttgt ggagcatggc
     1081 ctgtttgtca agcactgcaa agtggaagtg tatttgctgg agctgaagct ctgtgagaac
     1141 agtgacccca ccaatgtgct aagttgccat tttagcaaag cagacaccat tgcaactatt
     1201 gagaaggaga tgaggaagct cttcaacatc cctgcagaac gtgaaacacg gctttggaac
     1261 aaatacatga gcaacaccta tgagcagttg agcaagctag acaacactat ccaggatgct
     1321 gggctgtacc agggtcaggt gctagtaatt gagccccaaa atgaagatgg cacatggccc
     1381 cggcagagcc tgcagtcaaa atcaagcact gcacctagca gaaatttcac tacctcttca
     1441 aaaccatccg caagtcccta ttgctcagtg tctgcctctc tcattgcaaa tggtgatagc
     1501 actaacagct ctgggatgca cagctccggt gtcagcaggg gtggatctgg cttctctgct
     1561 tcgtataatt gccaggagcc cccatcacct catatacagc ctggcctctg tggacttgga
     1621 aacctgggga acacttgctt catgaactct gctctgcagt gcctgagcaa cactgcccca
     1681 ctgactgagt actttctcaa agatgagtat gaggccgaga tcaaccgaga caaccctctg
     1741 gggatgaaag gggagattgc agaggcctat gcagagctca tcaagcagat gtggtctgga
     1801 agggacactc acgtggcacc acggatgttc aagacgcaag tgggacgttt tgcccctcag
     1861 ttttctggct accaacagca agactcccag gagctgctag cctttattct agacggactg
     1921 cacgaggacc tgaaccgcgt aaagaagaag ccttacctgg agcccaagga cgccaatggg
     1981 cgaccagatg cggtggtagc aaaggaagcc tgggaaaacc acaggctgag gaatgattct
     2041 gtgattgtgg atactttcca tggccttttc aaatcgactt tggtttgccc agaatgtgct
     2101 aaagtttctg tgacctttga cccattttgc tatctaactc tcccactgcc tttgaagaag
     2161 gatcggatta tggaggtctt cctggttcct gctgaccctc agtgcagacc tatccagtac
     2221 cgtgtgactg tgccattgat gggggccatt tctgacctgt gtgaagcact ctccaagctg
     2281 tctggcattg ctgcagaaaa tatggtggtc actgatgtat ataatcaccg cttccacaaa
     2341 attttccaaa tggatgaagg tttaagccac atcacgcctc gagatgacat ttttgtgtat
     2401 gaggtctgca acacgtccat ggatggctca gagtgtatca ctcttccagt ctacttcaga
     2461 gagaagaagt ccaggccgtc gagtgcttcc tcgggggctg tgctctatgg acagcccctt
     2521 cttgtctctg tccctaagca taagctaacc ctagagtctt tgtaccaggc tgtttgtgat
     2581 cgtatcagcc gctacataaa acagcctttg cctgatgagt ttctcagctc acccttagag
     2641 cctggggcct gcaatggctc taggagtagc tatgaaggag atgaagagga agaaatggat
     2701 catcaagagg aaggaaaaga gcagctttcc gaagtggaag gcagtggtga ggacgatcag
     2761 ggagatgacc atagcgagag cgcccaaaag gtgaaaggcc agccgaggca caagaggctc
     2821 tttaccttca gcctcgtgaa ctcctgtgga actgctgata tcaattcact ggcaactgac
     2881 gggaaactcc tcaaactcaa ctctcgatcc acactggcca ttgactggga cagtgagacc
     2941 cgaagccttt actttgatga gcaagaatct gaggcctgtg agaagcacct gagcatgtca
     3001 cagccgcaga agaagaagaa ggcggcagta gccctgcgag agtgcatcga gctcttcacc
     3061 accatggaga cccttgggga gcatgacccc tggtactgtc ccacctgtaa gaagcaccag
     3121 caggcaacaa agaagtttga cttgtggtct ttgcccaaga tcctggtggt tcacctcaag
     3181 cgtttctcct ataacagata ctggcgggat aaacttgaca ccgtggtgga gttcccagtc
     3241 agagctctga acatgtccga gtttgtctgt gaccggtcag caaggcctta tgtttatgac
     3301 ctaattgctg tgtccaatca ctatggagcc atgggggttg gtcactacac tgcatatgcg
     3361 aagaacagac tgaacgggaa atggtattac tttgatgata gcagtgtgtc cctggcctct
     3421 gaggaccaga tagtgacgaa agccgcctac gtgttgttct atcagcgtcg ggatgacgaa
     3481 tgctccagca catcttcgct cggcagtttc cccggttctg atggaggggt gaagctgagc
     3541 agctcacacc agggcatggg ggatgaggag gcttacaaca tggacaccaa cctcgaggaa
     3601 caaaaactaa tatcagagga agatttggaa cagaaactaa tcagtgaaga agatttagag
     3661 caaaagttaa tttccgagga ggacttatag tgagggcccg atatctcgac aatcaacctc
     3721 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3781 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3841 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3901 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3961 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     4021 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4081 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     4141 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     4201 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4261 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac
     4321 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4381 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4441 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4501 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4561 cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc
     4621 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4681 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4741 ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt
     4801 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4861 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4921 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4981 ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5041 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5221 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5281 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5341 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5401 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5461 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5521 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5581 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5641 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5701 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5761 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5821 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5881 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5941 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     6001 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6061 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6121 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6181 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6241 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     6301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     6481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     6541 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     6601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     6661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     6721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     6781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     7081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     7141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     8041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     8101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     8161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     8761 gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
     8821 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     8881 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
