LOCUS       V020883                 9845 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020883
VERSION     V020883
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..4525
                     /label="TTLL5(NM_015072)"
                     /note="TTLL5(NM_015072)"
                     /gene="TTLL5"
     CDS             4532..4597
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4621..5209
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5275..5323
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5525..5953
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5967..6296
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6363..7157
                     /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    7333..7454
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7503..7519)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7527..7543
                     /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(7551..7581)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7596..7617
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7905..8493)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8664..9524)
                     /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(9525..9629)
                     /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 ccatgccaat cgtgatggcc cgggacctgg aggaaacagc
      721 atcatcctca gaggatgagg aggtcataag tcaagaggat catccatgca tcatgtggac
      781 tggaggctgc aggagaattc cagttttggt attccatgcc gacgctattc ttacaaagga
      841 caacaatatt agagtaattg gagaacgtta tcatttgtct tataagattg tacgaacgga
      901 cagtcgccta gtacgcagca ttctgacagc ccatggattt catgaagttc acccaagcag
      961 cactgactat aacctaatgt ggacaggatc ccacctgaag cccttcttac tgcgcaccct
     1021 ctctgaagca caaaaagtta atcactttcc caggtcttat gaacttaccc ggaaggaccg
     1081 actgtacaaa aacattattc gaatgcagca tacacatgga ttcaaggctt ttcacatcct
     1141 cccccagacc ttcctcctgc cagctgagta cgcggaattt tgtaattcat attcgaagga
     1201 ccggggacct tggatagtaa aaccagtggc atcttcaagg gggcggggcg tctacctgat
     1261 caacaatcca aaccagatct ccctggaaga gaacattttg gtctcccgtt acattaacaa
     1321 ccccctgctc atagatgatt tcaagtttga cgtgcgcctc tatgtgctcg tgacttccta
     1381 tgatcctctt gtcatctatc tctatgaaga aggattggct aggtttgcaa ctgtgcgata
     1441 tgatcaagga gccaagaaca ttcggaacca gttcatgcat ctgacaaact acagtgtcaa
     1501 caagaaaagt ggagattacg tcagttgtga cgatccagaa gtggaggatt atggaaacaa
     1561 atggagcatg agtgctatgc ttaggtacct gaaacaagaa ggcagagata caaccgcatt
     1621 gatggcccat gtagaagacc tgatcattaa gactataatc tctgctgaac tagctattgc
     1681 tactgcctgt aaaacctttg ttcctcatcg cagcagttgt tttgaactct atggctttga
     1741 cgtgctcata gattctactc tgaagccatg gttgttggaa gtgaatctct ctccttcttt
     1801 ggcctgtgat gcgcctctgg acctaaagat taaagccagt atgatttcag atatgttcac
     1861 tgttgtagga tttgtgtgcc aagatcctgc ccagcgggca tcaactcggc caatttatcc
     1921 cacctttgag tcttccaggc gaaacccttt ccagaaacct cagcgttgcc gtccactctc
     1981 tgccagtgat gcggaaatga aaaacctcgt gggctcagcc cgggagaaag ggccagggaa
     2041 gttgggtggt tctgtgcttg gtctgtcaat ggaggagatc aaagttttac gaagggtgaa
     2101 ggaggagaat gatcggcgag gtggatttat tcgcatattt cctacatctg agacatggga
     2161 aatatatggg tcctacctcg agcataagac ctcaatgaac tatatgctgg caacacgcct
     2221 cttccaggac agaatgactg ctgatggagc gccagaattg aagatagaga gtctgaattc
     2281 aaaggccaag ctgcatgctg cactttacga gaggaagctc ctgtctctgg aggtgcgaaa
     2341 acgtagacga cggagtagca gattgagggc aatgaggcca aaatacccag tgattaccca
     2401 accagctgaa atgaatgtta aaactgagac agagagtgaa gaggaggaag aagtcgcatt
     2461 agataatgaa gatgaagaac aggaggcttc ccaggaggag tctgcaggat ttcttagaga
     2521 aaatcaagcc aaatatacac cctcattgac agctttggta gaaaatacac ccaaagaaaa
     2581 ttccatgaaa gttcgtgaat ggaataataa aggtggacac tgctgcaaac ttgagactca
     2641 ggagctagag cctaaattta acctgatgca gattcttcaa gataatggca atcttagcaa
     2701 aatgcaggcc cgaatagcat tctctgccta tctccagcat gttcaaattc gcctgatgaa
     2761 agacagtggc ggtcagacgt tcagtgccag ttgggctgcc aaagaggatg aacagatgga
     2821 gctggttgtt cgtttcctca agcgagcatc aaataacctc cagcattcac tgaggatggt
     2881 attacccagt cgacgattgg cacttctgga acgcagaaga atcctggccc accagctggg
     2941 tgactttatc attgtataca acaaggaaac agaacaaatg gctgaaaaga aatcaaagaa
     3001 gaaagttgag gaagaagagg aagatggggt gaatatggaa aactttcagg agttcatcag
     3061 acaagcaagt gaggctgaac tggaggaggt gttgactttt tatacccaaa agaacaagtc
     3121 tgctagtgtc ttcctgggga ctcactctaa aatttctaag aacaacaaca attattctga
     3181 tagtggggca aaaggtgatc accctgagac tataatggaa gaagtgaaaa taaagccacc
     3241 taaacagcaa cagacgacag aaattcattc tgataaatta tctcgattta ccacttcagc
     3301 agaaaaagag gcaaaattag tttatagcaa ttcctcctct ggtcctactg ctactctgca
     3361 gaaaattccc aacacccatt tgtcatctgt tacaacctct gacctctctc cagggccttg
     3421 ccaccattct tctttatctc aaattccttc agctatcccc agcatgcctc accagccaac
     3481 aattttactg aacacagtct ctgccagtgc ttctccctgc ctacatcccg gggcacagaa
     3541 catcccaagc cctactggcc tgccacgctg tcgatcagga agtcacacca ttggtccctt
     3601 ttcttccttc caaagtgctg cacacatcta tagccagaaa ctgtctcgtc cctcttcagc
     3661 aaaggcagga tcgtgctatc taaacaagca tcattcagga atagccaaaa cacaaaaaga
     3721 gggagaagat gcttctttat atagcaaacg gtacaaccaa agtatggtta cagctgaact
     3781 tcagcggcta gctgagaagc aggcagcgag acagtattct ccatccagcc acatcaacct
     3841 cctcacccaa caggtaacaa acctgaattt ggcaactggc atcataaaca gaagcagtgc
     3901 ttcagctccc ccaaccctcc gacccatcat cagtcctagt ggcccgacat ggtctacaca
     3961 gtcagacccc caagctcccg agaatcactc cagctctcct ggaagcagga gcctgcagac
     4021 agggggattt gcctgggaag gagaagtaga aaacaacgtg tacagccagg ctacaggggt
     4081 ggtcccccag cacaagtatc accccacagc aggcagctat cagcttcaat ttgccctgca
     4141 gcaacttgaa caacaaaaac ttcagtcccg gcagctcctg gaccagagtc gagcccggca
     4201 ccaggcaatc tttggcagcc agacactacc taactccaat ttatggacaa tgaataatgg
     4261 tgcaggttgt agaatttcca gtgccacagc tagtggccag aagccaacca ctctgccaca
     4321 aaaagtggta ccacctccaa gttcttgcgc ctccctggtt cccaaacccc cacccaacca
     4381 cgaacaagtg ctcagaaggg caacatccca gaaagcttcc aaagggtcct ccgcggaagg
     4441 gcagctgaat ggactccaga gcagccttaa ccctgcagcc tttgtgccca tcaccagctc
     4501 tacagatcct gctcacacta aaatactcga ggactacaaa gaccatgacg gtgattataa
     4561 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac
     4621 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4681 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4741 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4801 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4861 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4921 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4981 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     5041 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     5101 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     5161 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     5221 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     5281 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5341 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5401 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5461 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     5521 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     5701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5881 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5941 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     6001 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     6061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6121 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     6181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     6241 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     6301 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     6361 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     6421 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6481 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6541 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6601 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6661 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6721 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6781 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6841 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6901 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6961 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     7021 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     7081 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     7141 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     7201 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     7261 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     7321 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7381 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7441 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     7501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     7681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     8041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     8101 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     8161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     8221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     8341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     8521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     9001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     9061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     9121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     9181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     9241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     9301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     9361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     9421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     9481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     9661 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     9721 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     9781 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     9841 ttagg
//
