LOCUS       V033987                 9178 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033987
VERSION     V033987
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..4668
                     /label="SMC1B(NM_148674)"
                     /note="SMC1B(NM_148674)"
                     /gene="SMC1B"
     CDS             4675..4740
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4778..5002
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5048..5476
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5490..5819
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5886..6680
                     /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    6854..6975
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7024..7040)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7048..7064
                     /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(7072..7102)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7117..7138
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7426..8011)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8182..9042)
                     /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(9043..9147)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggccc acctggagct gctgcttgtg gaaaatttca agtcgtggcg gggccgccag
     1021 gtcattggcc ccttccggag gttcacctgc atcatcggcc ccaacggctc tggaaaatct
     1081 aatgtaatgg atgcacttag ttttgtaatg ggagagaaaa tagctaattt aagagtgaaa
     1141 aatattcaag aactcattca tggagcacat attggaaaac ctatttcttc ttctgcaagt
     1201 gtaaaaatta tatatgtgga ggaaagtggc gaagagaaaa catttgcaag gattatccga
     1261 gggggatgct cagaatttcg ctttaatgat aatcttgtga gtcgttctgt ttacattgca
     1321 gagttggaaa agataggcat aatagtcaaa gcacaaaatt gtttggtttt tcagggaact
     1381 gtagagtcaa tttcagtgaa gaaacccaaa gaaaggaccc agttttttga ggaaatcagc
     1441 acttcaggag agcttatagg agaatatgaa gaaaagaaaa gaaagttaca aaaagccgaa
     1501 gaggatgcac agtttaactt taataagaaa aaaaatatag cggcagagcg cagacaagca
     1561 aaattagaga aggaagaggc agaacgttac cagagtctcc ttgaagaact gaaaatgaac
     1621 aagatacaac tgcagctttt tcaactatac cataatgaga aaaagattca tctcctgaac
     1681 accaagttag agcatgtgaa tagggatttg agtgtcaaaa gagagtcttt gtctcatcat
     1741 gaaaacatag ttaaagccag gaaaaaggaa catggaatgc taactagaca actacaacaa
     1801 acagaaaaag aattaaaatc ggttgaaacc cttttaaatc agaagaggcc tcagtacatt
     1861 aaagccaaag aaaacacttc tcaccacctt aagaaattag atgtggctaa gaaatcaata
     1921 aaggacagcg aaaaacaatg ttctaaacag gaagatgata taaaagccct ggagacagag
     1981 ctggctgatt tagatgctgc atggagaagt tttgaaaagc agattgagga agaaatttta
     2041 cataaaaagc gagacattga actggaagcc agtcagctgg atcgttataa agaacttaag
     2101 gaacaagtaa gaaagaaagt agctacaatg actcaacaac tggaaaaact gcagtgggaa
     2161 cagaagacag atgaagaaag actggcattt gaaaagagga ggcatggaga agttcaggga
     2221 aatctaaaac aaataaaaga acaaatagaa gatcataaaa aacgaataga gaagttagag
     2281 gagtatacaa agacatgcat ggattgcttg aaagagaaaa aacagcaaga ggaaacccta
     2341 gtggatgaaa ttgaaaaaac aaaatcaaga atgtctgaag ttaatgaaga attgaatctt
     2401 attagaagtg aattgcagaa tgctgggatt gatacccatg agggaaaacg tcagcaaaag
     2461 agagcagagg ttctggaaca ccttaaaaga ctgtacccag attctgtgtt tggaagacta
     2521 tttgacctgt gtcatcctat tcataagaaa taccagctgg ctgttactaa ggtttttggc
     2581 cggttcatca ctgccattgt tgtagcctct gaaaaggtag caaaagattg tattcgattt
     2641 ctgaaggagg aaagagctga acctgagaca ttcctcgctc tagattacct tgatatcaag
     2701 ccaatcaatg aaagactaag ggagcttaaa ggctgtaaaa tggtgattga tgtcataaag
     2761 actcagtttc ctcagctgaa gaaagtgatt cagtttgtgt gtggaaatgg tcttgtttgt
     2821 gagactatgg aagaagcaag gcatattgca ctcagtggac ctgaaagaca gaaaacagta
     2881 gctcttgatg gaacattatt tttaaaatct ggagtgatct ctggagggtc aagtgactta
     2941 aaatacaagg ctagatgctg ggatgagaaa gagttaaaga atctaagaga cagacgaagc
     3001 cagaaaatcc aagagctaaa gggtttaatg aagacactcc gcaaagaaac agatttgaaa
     3061 caaatacaga ccctgataca gggaactcaa acacgactca aatattcaca aaatgaacta
     3121 gagatgatta agaagaagca ccttgttgct ttttaccagg aacaatctca gttacaaagt
     3181 gaactactaa atattgagtc tcaatgtatt atgttgagtg aaggaatcaa ggaacgacaa
     3241 cgaagaatta aagaatttca agaaaagata gataaggtag aagacgatat cttccaacac
     3301 ttctgtgaag aaattggcgt ggaaaatatt cgtgaatttg agaacaaaca tgttaaacgg
     3361 caacaagaaa ttgatcaaaa aagattagaa tttgaaaaac aaaaaactcg gcttaatgtt
     3421 caacttgagt atagtcgcag tcaccttaag aagaaactga ataagatcaa cacattaaaa
     3481 gaaactatcc agaaaggtag tgaagatatt gatcacctaa agaaggctga agaaaactgt
     3541 ctgcagacag tgaatgaact catggcaaag cagcagcaac ttaaggacat acgtgtcact
     3601 cagaactcca gtgccgagaa agttcaaact caaattgaag aggaacggaa gaagtttctg
     3661 gctgttgata gggaagtggg gaaattgcaa aaagaagttg taagtattca aacttctctg
     3721 gaacagaaac gattagagaa gcataacttg ctgcttgatt gcaaagtgca agacattgag
     3781 ataatccttt tgtcggggtc actggatgac atcattgaag tggagatggg aactgaagca
     3841 gaaagtaccc aggcaacaat tgatatctat gaaaaagaag aagcctttga aatagactac
     3901 agctctctaa aagaggattt gaaggctcta cagtctgatc aagaaatcga ggcccacctt
     3961 aggctcttat tgcagcaagt agcatcccag gaagatatct tactgaaaac agcagcccca
     4021 aacctacgag cactggagaa cttaaagact gtcagagaca agtttcaaga gtccacagat
     4081 gcttttgagg ccagcagaaa ggaagccaga ctgtgtaggc aagagttcga gcaagtgaaa
     4141 aaaaggagat acgatctttt cacccagtgt tttgagcatg tctcaatctc aattgatcaa
     4201 atctacaaga agctctgcag aaacaacagc gcccaagcat ttcttagccc agagaaccct
     4261 gaagaacctt acttggaggg aattagctat aactgtgtgg ccccaggcaa acggtttatg
     4321 ccaatggaca atttgtcagg gggagaaaag tgtgtggcag ccttggctct cctgtttgct
     4381 gtgcacagtt ttcgtcctgc cccattcttt gttttagatg aagtggatgc agccctagac
     4441 aatactaaca taggcaaagt gtcaagttac atcaaagagc aaactcaaga ccagtttcag
     4501 atgatagtca tctccctaaa agaagagttc tattccagag ccgacgcgct gatcggcatc
     4561 tatcctgagt acgatgactg catgttcagc cgagttttga ccctagatct ttctcagtat
     4621 ccagacactg aaggccaaga aagcagcaag agacacggag agtcccgcct cgaggactac
     4681 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     4741 tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4801 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4861 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4921 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4981 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     5041 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5101 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5161 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5221 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5281 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5341 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5401 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5461 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5521 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5641 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5701 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5761 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5821 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5881 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5941 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6001 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6061 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6121 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6181 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6241 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6301 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6361 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6421 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6481 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6541 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6601 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6661 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6721 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6781 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6841 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6901 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6961 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7021 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7081 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7141 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     7981 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8041 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8101 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8161 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8221 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8281 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8341 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8401 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8461 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8521 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8581 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8641 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8701 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8761 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8821 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8881 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8941 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9001 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9061 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9121 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
