LOCUS       V020755                 8675 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020755
VERSION     V020755
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..3355
                     /label="TTLL6(NM_001130918)"
                     /note="TTLL6(NM_001130918)"
                     /gene="TTLL6"
     CDS             3362..3427
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3451..4039
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4105..4153
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4355..4783
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4797..5126
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5193..5987
                     /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    6163..6284
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6333..6349)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6357..6373
                     /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(6381..6411)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6426..6447
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6735..7323)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7494..8354)
                     /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(8355..8459)
                     /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 ccatgggagc gttactcctt catccttcga ggagggggcc
      721 ggcaggtgtg gttgcaagct ggactagcag cccagcgggg cgagacgggg gagtaggaat
      781 tgcgggggcc tggtatttcc ccagagcctc ctcccaggcc agggagatgc cacagtgccc
      841 gactttggaa agccaggaag gggaaaactc cgaagagaag ggggacagtt ccaaagaaga
      901 tccaaaagaa accgtcgcgc tggcttttgt gagagagaac ccaggggcac aaaacggact
      961 tcagaatgcc cagcagcaag gcaagaagaa gaggaagaaa aagagattgg tgatcaatct
     1021 atccagctgc cggtatgaga gtgtgcgcag ggctgcccaa cagtacggct ttagagaggg
     1081 aggggaagac gatgactgga ctctctattg gacagattac tcagtgtcac tggagcgggt
     1141 gatggaaatg aaaagttacc agaagatcaa tcacttcccc gggatgagtg aaatctgccg
     1201 gaaggacttg ctggccagga acatgagccg catgttaaag atgttcccta aagatttccg
     1261 ctttttccct aggacctggt gtcttcctgc tgactgggga gatttgcaga cctacagcag
     1321 gtcaagaaaa aataagacat acatttgtaa gccggattcg ggctgccaag ggaaaggtat
     1381 attcatcacc cggacagtga aagaaatcaa accaggggag gatatgatct gtcagctgta
     1441 tatttcaaag ccctttatca ttgatgggtt taagtttgac ctacggattt atgtactggt
     1501 gacatcctgt gaccctctca ggatttttgt gtacaatgaa ggactggccc gctttgcgac
     1561 gacctcttac tcccgccctt gcacagacaa cctggatgat atctgcatgc acctgactaa
     1621 ttattccatt aataagcaca gttcaaattt cagtcgagat gcacactctg gcagtaagag
     1681 gaagctctcc accttcagtg catacttgga ggaccacagc tacaacgtgg agcagatatg
     1741 gagggatatt gaggacgtca tcatcaagac cctcatctcg gcccacccca tcatcaggca
     1801 taactaccac acctgcttcc ccaaccacac actcaacagc gcctgctttg agatcctggg
     1861 ctttgacatt ttgttggacc acaaactcaa accctggctg ctggaggtca accactctcc
     1921 aagcttctcc accgactctc ggttggataa agaggtgaaa gatggtctgc tgtatgacac
     1981 cttagtcctg atcaacctgg aaagctgtga caagaagaaa gtcttggagg aggagagaca
     2041 acgggggcag ttcctgcagc agtgttgttc tcgggagatg aggattgagg aagccaaggg
     2101 tttccgggcc gtgcagttaa agaaaactga aacgtatgag aaggaaaact gtggagggtt
     2161 ccgactgatt tatcccagtc tgaattcgga gaagtatgag aagtttttcc aggacaacaa
     2221 ctccctcttc cagaatactg ttgcttccag ggctcgggag gagtatgccc ggcaactgat
     2281 ccaggagctg agactaaaac gggagaaaaa gcccttccaa atgaagaaga aggtagagat
     2341 gcagggggaa tcggcaggcg agcaagtgag aaagaagggc atgaggggct ggcaacagaa
     2401 acaacagcag aaagacaagg ccgccaccca agcctccaaa cagtacatcc agccattgac
     2461 attagtatcc tacacacctg acttgctctt gagtgtcaga ggtgaaagga aaaatgaaac
     2521 agacagcagc ctcaaccagg aggctcccac ggaggaggcc agctctgttt tccccaagct
     2581 gacgtctgcg aagcccttca gttctctacc cgatctgagg aatatcaatc tcagcagctc
     2641 gaagttggag cccagtaaac ccaacttcag catcaaggag gccaagtctg cctctgcagt
     2701 gaacgtattc actggcactg tgcacttaac ctccgtagaa accaccccag aatccaccac
     2761 ccaactctca atctccccaa agtctccgcc aaccctggct gtgaccgcca gctctgagta
     2821 cagtggccca gagacggaca gggtggtatc ctttaaatgc aagaagcagc agacccctcc
     2881 acacttaacc cagaagaaaa tgttaaaatc ttttctgccc acaaaatcca agagcttctg
     2941 ggagagtccg aacacaaact ggactttgct aaagagtgac atgaacaagc cacatttgat
     3001 atccgagcta ctcaccaagc ttcaactgag tgggaagctc tccttcttcc cagctcacta
     3061 caaccccaag ctggggatga ataacctgtc acaaaacccc tccctgcctg gggagtgcca
     3121 ctcccgcagt gacagctctg gcgagaagag gcagctggat gtgtcctccc tcctcttgca
     3181 gagtcctcag agctataatg ttactctgag ggacctgctg gtgattgcca ctccagccca
     3241 actggatcca aggccttgta gaagccacgc aagtgctatg agggacccat gtatgcagga
     3301 tcaagaagca tacagccatt gcctgatctc tggccaaaaa ggatgtgaga ggagcctcga
     3361 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     3421 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3481 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3541 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3601 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3661 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3721 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3781 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3841 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3901 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3961 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4021 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4081 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4141 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4201 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4261 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4321 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4381 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4441 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4501 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4561 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4621 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4681 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4741 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4801 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4861 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4921 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4981 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5041 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5101 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5161 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5221 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5281 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5341 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5401 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5461 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5521 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5581 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5641 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5701 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5761 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5821 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5881 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5941 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6001 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6061 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6121 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6181 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6241 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6301 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6361 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6421 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6541 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6901 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7141 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8341 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8401 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8461 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8521 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8581 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8641 gcttgaccga caattgcatg aagaatctgc ttagg
//
