LOCUS       V042888                 8545 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042888
VERSION     V042888
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..3201
                     /label="KAT2B(NM_003884)"
                     /note="KAT2B(NM_003884)"
                     /gene="KAT2B"
     CDS             3208..3297
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3321..3909
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3975..4023
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4225..4653
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4667..4996
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5063..5857
                     /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    6033..6154
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6203..6219)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6227..6243
                     /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(6251..6281)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6296..6317
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6605..7193)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7364..8224)
                     /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(8225..8329)
                     /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 ccaccatgtc cgaggctggc
      721 ggggccgggc cgggcggctg cggggcagga gccggggcag gggccgggcc cggggcgctg
      781 cccccgcagc ctgcggcgct tccgcccgcg cccccgcagg gctccccctg cgccgctgcc
      841 gccgggggct cgggcgcctg cggtccggcg acggcagtgg ctgcagcggg cacggccgaa
      901 ggaccgggag gcggtggctc ggcccgaatc gccgtgaaga aagcgcaact acgctccgct
      961 ccgcgggcca agaaactgga gaaactcgga gtgtactccg cctgcaaggc cgaggagtct
     1021 tgtaaatgta atggctggaa aaaccctaac ccctcaccca ctccccccag agccgacctg
     1081 cagcaaataa ttgtcagtct aacagaatcc tgtcggagtt gtagccatgc cctagctgct
     1141 catgtttccc acctggagaa tgtgtcagag gaagaaatga acagactcct gggaatagta
     1201 ttggatgtgg aatatctctt tacctgtgtc cacaaggaag aagatgcaga taccaaacaa
     1261 gtttatttct atctatttaa gctcttgaga aagtctattt tacaaagagg aaaacctgtg
     1321 gttgaaggct ctttggaaaa gaaaccccca tttgaaaaac ctagcattga acagggtgtg
     1381 aataactttg tgcagtacaa atttagtcac ctgccagcaa aagaaaggca aacaatagtt
     1441 gagttggcaa aaatgttcct aaaccgcatc aactattggc atctggaggc accatctcaa
     1501 cgaagactgc gatctcccaa tgatgatatt tctggataca aagagaacta cacaaggtgg
     1561 ctgtgttact gcaacgtgcc acagttctgc gacagtctac ctcggtacga aaccacacag
     1621 gtgtttggga gaacattgct tcgctcggtc ttcactgtta tgaggcgaca actcctggaa
     1681 caagcaagac aggaaaaaga taaactgcct cttgaaaaac gaactctaat cctcactcat
     1741 ttcccaaaat ttctgtccat gctagaagaa gaagtatata gtcaaaactc tcccatctgg
     1801 gatcaggatt ttctctcagc ctcttccaga accagccagc taggcatcca aacagttatc
     1861 aatccacctc ctgtggctgg gacaatttca tacaattcaa cctcatcttc ccttgagcag
     1921 ccaaacgcag ggagcagcag tcctgcctgc aaagcctctt ctggacttga ggcaaaccca
     1981 ggagaaaaga ggaaaatgac tgattctcat gttctggagg aggccaagaa accccgagtt
     2041 atgggggata ttccgatgga attaatcaac gaggttatgt ctaccatcac ggaccctgca
     2101 gcaatgcttg gaccagagac caattttctg tcagcacact cggccaggga tgaggcggca
     2161 aggttggaag agcgcagggg tgtaattgaa tttcacgtgg ttggcaattc cctcaaccag
     2221 aaaccaaaca agaagatcct gatgtggctg gttggcctac agaacgtttt ctcccaccag
     2281 ctgccccgaa tgccaaaaga atacatcaca cggctcgtct ttgacccgaa acacaaaacc
     2341 cttgctttaa ttaaagatgg ccgtgttatt ggtggtatct gtttccgtat gttcccatct
     2401 caaggattca cagagattgt cttctgtgct gtaacctcaa atgagcaagt caagggctat
     2461 ggaacacacc tgatgaatca tttgaaagaa tatcacataa agcatgacat cctgaacttc
     2521 ctcacatatg cagatgaata tgcaattgga tactttaaga aacagggttt ctccaaagaa
     2581 attaaaatac ctaaaaccaa atatgttggc tatatcaagg attatgaagg agccacttta
     2641 atgggatgtg agctaaatcc acggatcccg tacacagaat tttctgtcat cattaaaaag
     2701 cagaaggaga taattaaaaa actgattgaa agaaaacagg cacaaattcg aaaagtttac
     2761 cctggacttt catgttttaa agatggagtt cgacagattc ctatagaaag cattcctgga
     2821 attagagaga caggctggaa accgagtgga aaagagaaaa gtaaagagcc cagagaccct
     2881 gaccagcttt acagcacgct caagagcatc ctccagcagg tgaagagcca tcaaagcgct
     2941 tggcccttca tggaacctgt gaagagaaca gaagctccag gatattatga agttataagg
     3001 ttccccatgg atctgaaaac catgagtgaa cgcctcaaga ataggtacta cgtgtctaag
     3061 aaattattca tggcagactt acagcgagtc tttaccaatt gcaaagagta caacccccct
     3121 gagagtgaat actacaaatg tgccaatatc ctggagaaat tcttcttcag taaaattaag
     3181 gaagctggat taattgacaa gctcgaggaa caaaaactaa tatcagagga agatttggaa
     3241 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatag
     3301 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3361 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3421 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3481 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3541 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3601 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3661 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3721 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3781 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3841 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3901 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3961 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     4021 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4081 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4141 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     4201 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4261 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4321 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4381 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4441 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4501 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4561 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     4621 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4681 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4741 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4801 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4861 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4921 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4981 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     5041 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5101 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5161 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5221 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5281 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5341 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5401 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5461 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5521 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5581 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5641 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5701 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5761 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5821 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     5881 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5941 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6001 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6061 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6121 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6181 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6781 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7021 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7141 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7201 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7261 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7321 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7441 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7561 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7801 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7861 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8041 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8221 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8281 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8341 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     8401 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     8461 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     8521 caattgcatg aagaatctgc ttagg
//
