LOCUS       V020705                 7319 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020705
VERSION     V020705
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..1999
                     /label="TTLL9(NM_001008409)"
                     /note="TTLL9(NM_001008409)"
                     /gene="TTLL9"
     CDS             2006..2071
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2095..2683
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2749..2797
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2999..3427
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3441..3770
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3837..4631
                     /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    4807..4928
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4977..4993)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5001..5017
                     /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(5025..5055)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5070..5091
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5379..5967)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6138..6998)
                     /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(6999..7103)
                     /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 ccatggtgcc atccagggaa gctctgctgg gaccaggcac
      721 aactgccatt aggtgcccca agaaattaca gaaccaaaat tacaagggcc atggattgtc
      781 aaagggaaaa gagcgagagc agagagcatc gatccggttc aagaccaccc tcatgaacac
      841 actcatggac gtccttcgcc acaggccagg atgggtggaa gtgaaggacg aaggggagtg
      901 ggatttctac tggtgtgacg tcagctggct ccgggagaac ttcgaccaca cctacatgga
      961 tgaacatgtg cggatcagtc acttccggaa ccactatgag ctgacccgga agaactacat
     1021 ggtgaagaac ctgaagcggt tccggaagca gctggagcgt gaggcaggaa agctggaggc
     1081 agccaagtgt gacttcttcc ccaaaacctt tgagatgcct tgcgagtacc acctgtttgt
     1141 agaggagttt cgcaaaaacc caggaatcac ctggatcatg aagcctgtag cccggtctca
     1201 agggaaaggc atcttcctct tccgtaggct gaaggacatc gtggactgga ggaaggacac
     1261 aagaagctct gacgaccaga aagatgatat tcccgtggag aactatgtgg ctcagcgtta
     1321 cattgaaaat ccttacctga taggaggccg caagtttgac ctgcgtgtct atgtgctggt
     1381 gatgtcggtg tttgctgaat gcctgctgtg gtctgggcac aggagacagg atgttcacct
     1441 caccaacgtg gctgtgcaaa aaacatctcc cgactaccac ccaaagaagg gctgcaagtg
     1501 gacgctgcag cgcttccggc agtacctggc gtccaaacac gggcccgagg cagtggagac
     1561 actcttcagg gacatcgaca acatctttgt caaaagcctg cagagtgtgc agaaggtgat
     1621 catcagtgac aagcactgct tcgagctgta cggctatgac atcctcatcg accaggacct
     1681 caagccgtgg ctcctggagg tcaatgcgtc cccatcactg acagccagca gccaggaaga
     1741 ctatgagctc aagacctgcc tcctggaaga caccctgcat gttgtggaca tggaagcgag
     1801 gctcacggga agggagaagc gagtcggggg ctttgacctc atgtggaatg atggccctgt
     1861 tagcagagag gagggggctc ctgacctgtc gggaatggga aactttgtga ccaacacaca
     1921 tctcggctgc gtcaacgatc ggaagaaaca actgaggcag ctcttctgct cccttcaagt
     1981 tcagaagaaa gcttccagtc tcgaggacta caaagaccat gacggtgatt ataaagatca
     2041 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     2101 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2161 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2221 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2281 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2341 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2401 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2461 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     2521 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2581 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2641 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     2701 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     2761 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     2821 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     2881 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2941 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3001 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3061 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3121 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     3181 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3241 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3301 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3361 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3421 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3601 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3721 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3781 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3841 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3901 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3961 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4021 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4081 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4141 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4201 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4261 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4321 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4381 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4441 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4501 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4561 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4621 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     4681 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     4741 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     4801 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4861 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4921 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4981 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5041 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5101 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5161 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5221 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5281 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5341 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5401 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5461 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5521 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5581 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5641 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5701 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5761 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5821 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5881 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5941 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6001 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6061 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6121 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6181 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6241 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6301 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6361 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6421 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6481 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6541 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6601 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6661 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6721 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6781 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6841 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6901 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6961 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7021 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7081 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7141 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     7201 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     7261 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
