LOCUS       V019154                 8525 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019154
VERSION     V019154
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             683..3205
                     /label="BACH2(NM_021813)"
                     /note="BACH2(NM_021813)"
                     /gene="BACH2"
     CDS             3212..3277
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3301..3889
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3955..4003
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4205..4633
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4647..4976
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5043..5837
                     /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    6013..6134
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6183..6199)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6207..6223
                     /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(6231..6261)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6276..6297
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6585..7173)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7344..8204)
                     /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(8205..8309)
                     /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 ggatccgcca ccatgtctgt ggatgagaag cctgactccc ccatgtatgt
      721 gtatgagtcc acagtccact gcaccaacat cctcctgggc ctcaatgacc agcggaaaaa
      781 ggatattctc tgtgacgtga ctttgatcgt ggagaggaag gagttccggg cccaccgggc
      841 tgtgctggcc gcatgcagtg aatatttttg gcaggcgctg gttggacaga caaaaaatga
      901 tttggtggtc agcttgcctg aggaggtcac agccaggggc tttgggccgc tgttacagtt
      961 tgcctacact gccaagctgt tactcagcag agaaaacatc cgcgaggtca tccgctgtgc
     1021 tgagttcctg cgcatgcaca acctggagga ctcctgcttc agcttcctgc agacccagct
     1081 cctgaacagt gaggatggcc tgtttgtgtg ccggaaggat gctgcgtgcc agcgcccaca
     1141 cgaggactgc gagaactctg caggagagga ggaggatgaa gaggaggaga cgatggattc
     1201 agagacggcc aagatggctt gccccaggga ccagatgctt ccagagccca tcagctttga
     1261 ggccgccgcc atccccgtag cagagaagga agaagccctg ctgcccgagc ctgacgtgcc
     1321 cacagacacc aaggagagct cagaaaagga cgcgttaacg cagtacccca gatacaagaa
     1381 ataccagctt gcatgtacca agaatgtcta taatgcatca tcacacagta cctcaggttt
     1441 tgcaagcaca ttccgggaag ataactctag caacagcctc aagccggggc ttgccagggg
     1501 gcagattaaa agtgagccgc ccagtgaaga gaatgaggaa gagagcatca cgctctgcct
     1561 gtctggagat gagcctgacg ccaaggacag agcgggggat gtcgagatgg accggaaaca
     1621 gcccagccct gcccctaccc ccacggcccc agctggggcc gcctgcctgg agagatccag
     1681 gagcgtggcc tcgccctcct gcttaaggtc tctgttcagc ataacgaaaa gtgtggagct
     1741 gtctggcctg cccagtacat ctcagcagca ctttgccagg agtccagcct gcccttttga
     1801 caaggggatc actcagggtg accttaaaac tgactacacc cctttcacag ggaattatgg
     1861 acagccccac gtgggccaga aggaggtgtc caacttcacc atggggtcgc ccctcagggg
     1921 gcctgggttg gaggctctct gtaaacagga gggagagctg gaccggagga gcgtgatctt
     1981 ctcctccagc gcttgtgacc aagtgagcac ctcggtgcat tcttattctg gggtgagcag
     2041 tttggacaaa gacctctctg agccggtgcc aaagggtctg tgggtgggag ccggccagtc
     2101 cctccccagc tcgcaggcct actcccacgg tgggctgatg gccgaccact tgccaggaag
     2161 gatgcggccc aacaccagct gcccggtacc aatcaaagtc tgccctcgct cacccccctt
     2221 ggagaccagg accaggactt ccagctcctg ctcttcctat tcctacgcgg aggacgggag
     2281 cgggggctca ccctgcagcc tccctctctg tgagttctcc tcctcgccct gttcccaggg
     2341 agccagattc cttgccacag aacatcagga accaggcctg atgggagatg gaatgtacaa
     2401 ccaagtgcgg ccccaaatta aatgtgagca gtcttatgga accaactcca gtgacgaatc
     2461 cggatcgttc tcggaagcag acagtgagtc gtgtcctgtg caggacaggg gccaggaggt
     2521 aaaacttcct tttcctgtag atcaaatcac agatcttcca aggaacgatt tccagatgat
     2581 gattaaaatg cacaagctaa cctcagaaca gttagagttt attcatgatg tccgacggcg
     2641 cagcaagaac cgcatcgcgg cccagcgctg ccgcaaaagg aaactggact gtattcagaa
     2701 tttagaatgt gaaatccgca aattggtgtg tgagaaagag aaactgttgt cagagaggaa
     2761 tcaactgaaa gcatgcatgg gggaactgtt ggacaacttc tcctgccttt cccaggaagt
     2821 ttgccgagac atccagagcc ccgagcagat ccaggccctg catcggtatt gccctgtcct
     2881 cagacccatg gacttgccca cggcctccag tattaaccct gcgcccttgg gtgctgagca
     2941 gaacattgcg gcctcccaat gcgcagtggg ggaaaacgtg ccctgctgct tggagccagg
     3001 cgcggctccc cccggacccc cctgggcacc cagcaacacc tccgagaatt gtacctctgg
     3061 gaggagacta gaaggcactg acccgggaac cttctcagag agaggacctc ctcttgaacc
     3121 caggagccaa acagtgaccg tggacttctg ccaggaaatg actgataagt gtacaactga
     3181 cgaacagccc aggaaagatt ataccctcga ggactacaaa gaccatgacg gtgattataa
     3241 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     3301 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3361 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3421 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3481 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3541 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3601 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3661 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3721 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3781 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3841 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     3901 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3961 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4021 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4081 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4141 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4201 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4561 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4621 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4801 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4981 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5041 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5101 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5161 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5221 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5281 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5341 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5401 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5461 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5521 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5581 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5641 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5701 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5761 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5821 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5881 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5941 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6001 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6061 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6121 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6181 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6241 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6301 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6361 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6421 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6481 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6781 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7021 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7621 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8341 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     8401 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     8461 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8521 ttagg
//
