LOCUS       V026610                 8939 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026610
VERSION     V026610
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             941..4429
                     /label="AFF4(NM_014423)"
                     /note="AFF4(NM_014423)"
                     /gene="AFF4"
     CDS             4436..4501
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4539..4763
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4809..5237
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5251..5580
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5647..6441
                     /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    6615..6736
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6785..6801)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6809..6825
                     /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(6833..6863)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6878..6899
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7187..7772)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7943..8803)
                     /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(8804..8908)
                     /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 atccgccacc atgaaccgtg aagaccggaa
      961 tgtgctgcgt atgaaagaac gggaaaggcg gaatcaggaa attcagcagg gcgaagacgc
     1021 cttcccacct agctctcctc tctttgcaga gccatacaaa gttactagca aagaagataa
     1081 gttatcaagt cgtattcaga gtatgcttgg aaactacgat gaaatgaagg atttcatagg
     1141 agacagatct ataccaaagc ttgttgcaat tcccaagcct acagtaccac catcagcaga
     1201 tgaaaaatct aacccaaatt tctttgaaca gagacatgga ggctctcatc agagtagcaa
     1261 atggactcca gtaggacccg cacccagcac ttctcagtct cagaaacggt cctcaggctt
     1321 acagagtgga catagtagcc agcggaccag cgcaggtagc agtagtggca ctaacagtag
     1381 tggtcagagg cacgaccgtg agtcatataa caatagtggg agcagtagcc ggaaaaaagg
     1441 ccagcatgga tcagaacact ccaaatcacg ttcttccagc cctggaaaac cccaggctgt
     1501 ttcttcatta aactctagtc attccaggtc tcatgggaat gatcaccata gcaaggaaca
     1561 tcaacgctcc aaatcacctc gggaccctga tgcaaactgg gattctcctt cccgtgtacc
     1621 tttttcaagt gggcagcact caactcaatc tttcccaccc tcattgatgt caaagtccaa
     1681 ttcaatgtta cagaaaccca ctgcctatgt gcggcccatg gacggacagg agtccatgga
     1741 accaaagctg tcctctgagc actacagcag ccaatcccat ggcaacagca tgactgagct
     1801 gaagcccagc agcaaagcac atctcaccaa gctgaaaata ccttcccaac cactggatgc
     1861 atcagcttct ggtgatgtga gctgtgtgga tgaaatccta aaagagatga cgcattcatg
     1921 gcctccccct ctaacggcta ttcatacacc atgcaaaaca gaaccttcca aatttccttt
     1981 tccaactaag gagtctcagc agtccaattt tggcactgga gaacaaaaaa gatataatcc
     2041 ttctaaaact tcaaatgggc accagtctaa atctatgtta aaagatgact taaaactaag
     2101 cagcagtgaa gacagtgatg gggaacagga ttgtgataag acaatgccga ggagtacacc
     2161 aggaagtaac tctgaacctt cacaccataa tagtgaagga gcagataact ccagggatga
     2221 ttctagtagc cacagtggat ctgaaagcag ctctggatct gactcagaga gtgaaagtag
     2281 ttccagtgac agtgaggcaa atgagccatc ccagagtgca tctcccgagc ctgaaccccc
     2341 gccaacaaac aaatggcaac ttgataattg gctgaataaa gtgaacccac ataaagtgtc
     2401 acccgcctct tcagtggaca gtaacatccc atcatctcaa ggctacaaaa aggaaggccg
     2461 agagcagggc actgggaata gctacactga tacaagtgga cctaaagaaa cgagttccgc
     2521 tactccggga cgagactcca aaaccatcca aaagggatca gaaagtgggc gtgggaggca
     2581 gaaatctcct gcacagagtg acagcacaac acagagaaga actgtaggca aaaaacaacc
     2641 caaaaaggct gagaaggcag ctgctgaaga gcctcgtgga ggcctgaaga tagaaagtga
     2701 aacccctgta gacttggcta gcagcatgcc ctccagcaga cacaaagcag ccaccaaagg
     2761 ctcaaggaaa cccaatataa agaaggagtc taagtcttcc cctcgaccta cagcagagaa
     2821 aaagaaatat aagtcaacaa gtaaatcttc ccagaaatca agggaaatca tagaaacaga
     2881 tacctcatcc tcagattcag atgaaagtga gagccttcct ccttcctcac aaactcctaa
     2941 gtaccccgag agcaatagga ctcctgttaa accctcctca gtggaggaag aagatagctt
     3001 ttttcggcaa cgaatgttct ctcctatgga agagaaggaa cttctttcac ccctcagtga
     3061 gcctgatgac aggtacccac ttattgtgaa gattgacctg aatcttttga ctagaatacc
     3121 aggaaagcct tacaaagaaa cagagccgcc caagggggaa aagaaaaatg tgccagaaaa
     3181 gcacacgaga gaggctcaga aacaagcctc agaaaaagtt tccaacaaag gcaagaggaa
     3241 gcataagaat gaagatgata accgagccag tgagagcaag aaacccaaaa cggaggacaa
     3301 gaattcagca ggccataagc catccagcaa cagagagtca tctaagcaga gtgctgcaaa
     3361 agaaaaggat ttgttgcctt ctcccgctgg gcctgttcct tcaaaagatc caaaaacaga
     3421 gcatggctct cggaagagga ctattagtca gtcttcttcc ttaaagtcaa gcagtaacag
     3481 caacaaggag acgagtggca gcagcaaaaa cagttcctcc acatcaaagc agaagaagac
     3541 cgaagggaag acttccagta gctccaagga ggttaaggaa aaggctccaa gtagctcctc
     3601 taactgtcct ccatctgcac caactcttga ttcttctaag cctcggagaa caaagcttgt
     3661 ctttgatgac agaaattatt cagcagacca ttatttacaa gaagcaaaaa agctaaagca
     3721 caatgcagat gcattgtctg ataggtttga gaaagctgta tactatcttg atgctgtggt
     3781 atctttcatt gaatgtggga atgcattaga gaagaatgct caggaatcca aatccccatt
     3841 ccctatgtat tcagagacgg tggatctcat caaatacact atgaagctaa agaattactt
     3901 ggcaccagat gctacagctg cagataaacg actcacagta ctttgcctgc gatgcgagtc
     3961 tttgctgtac ctgaggctgt tcaaactgaa gaaggaaaat gctctgaagt actcaaagac
     4021 actgacagag cacctgaaga attcttataa taattctcaa gcaccatcgc ctggcttggg
     4081 aagcaaagct gtggggatgc cttcccctgt ttctccaaag ctgtcaccag gcaattcagg
     4141 aaattattca tctggggcca gtagtgcttc tgcaagtggt tcttcagtga ccattccaca
     4201 gaagatccac cagatggcag ccagctatgt tcaggtcaca tccaacttcc tctatgccac
     4261 cgaaatttgg gaccaagctg aacagctttc caaagagcaa aaagaattct ttgctgaact
     4321 ggataaagta atgggccctc tcatctttaa tgcaagcatc atgacagatc tagttcgtta
     4381 tacccggcag ggactgcact ggcttcgcca ggatgccaag ttgatatctc tcgaggacta
     4441 caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa
     4501 gtagtgaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     4561 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4621 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4681 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4741 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4801 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4861 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4921 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4981 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5041 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5101 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5161 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5221 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     5281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     5341 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5401 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5461 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     5521 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5581 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5641 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5701 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5761 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5821 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5881 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5941 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6001 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6061 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6121 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     6181 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6241 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6301 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6361 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6421 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6481 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6541 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6601 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6661 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6721 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     6781 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6841 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6901 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6961 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7021 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7081 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7141 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7201 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7261 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7321 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7381 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7441 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7501 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7561 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7621 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7681 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     7741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
