LOCUS       V037201                10059 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037201
VERSION     V037201
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4739
                     /label="Ttll5(NM_001081423)"
                     /note="Ttll5(NM_001081423)"
                     /gene="Ttll5"
     CDS             4746..4811
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4835..5423
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5489..5537
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5739..6167
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6181..6510
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6577..7371
                     /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    7547..7668
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7717..7733)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7741..7757
                     /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(7765..7795)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7810..7831
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8119..8707)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8878..9738)
                     /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(9739..9843)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc agttgtgatg gcccgggacc
      781 tggaggagac agcatcgtcc tcagaggacg aggacctggc caaccaagag gatcacccat
      841 gtattatgtg gactggaggc tgcaggagaa ttccagtttt ggttttccat gctgaggcta
      901 ttttgacaaa ggacaacaat attcgagtaa ttggagagcg ttatcatttg tcttataaga
      961 ttgtgagaac ggacagtcgc ctggtgcgaa gcattctgac agcccatgga tttcatgaag
     1021 ttcacccgag cagcactgac tataatttga tgtggacagg atcccaccta aaaccgttct
     1081 tacttcgaac cctctctgaa gcacaaaagg ttaatcattt ccccaggtct tatgaactaa
     1141 cccgcaagga ccgactgtac aaaaacatca ttcggatgca gcacactcat ggattcaaag
     1201 cattccacat cctccctcag acctttctcc tgccagctga gtatgcagaa ttctgtaatt
     1261 catattcaaa ggaccgagga ccttggatag taaaaccagt ggcctcttct agggggcggg
     1321 gcgtctactt gatcaacaac ccaaaccaga tctccctgga agagaacatc ttggtctccc
     1381 gttacattaa caaccccctg ctcatagatg atttcaagtt tgacgtgcgc ctctatgtgc
     1441 tcgtgacttc ctatgatccc cttgtcatct atctctatga ggaaggactg gctaggtttg
     1501 caactgtgcg ttatgatcaa ggctccaaga acatccgcaa ccagttcatg cacctgacga
     1561 actacagcgt gaacaagaag agtggagact atgtgagttg tgatgaccca gaagtggagg
     1621 attatgggaa caagtggagc atgagtgcca tgcttaggta cctgaaacaa gaaggcaagg
     1681 acacaactgc attgatggcc catgtggaag acctgatcat taagacaata atctctgctg
     1741 aactagctat cgccactgcc tgtaaaacct ttgttccaca ccgcagcagt tgttttgaac
     1801 tgtatggctt tgatgtgctc atagataata ctctgaagcc atggttgttg gaagtgaatc
     1861 tctctccttc tttggcctgt gatgctcctc tggatctaaa gattaaagcc agtatgatct
     1921 cagacatgtt cactgttgta ggatttgtgt gccaagatcc tgcccagcgg acatccaacc
     1981 ggtcaattta tcccagtttt gagtcttcca ggcggaaccc ttttcagaaa cctcagcgca
     2041 cccgtccact ctctgccagt gatgcagaaa tgaaaaacct tgtggcctca gcccgggaga
     2101 aggtaccggg gaagttaggc ggttctgtgc ttggtctgtc aatggaggag atcaaagttc
     2161 tccggagggt aaaggaggag aatgatcgga gaggagggtt tattcgcata tttcctacat
     2221 cggagacatg ggaaatatat gggtcctacc ttgaacacaa aacctccatg aactacatgc
     2281 tggcaacacg cctattccag gacagaggaa accccagaag gagcttgctg acaggaagag
     2341 cacgagtgag cacggaagga gcaccagagc taaaggtgga gagcatgaac tccaaggcca
     2401 agctgcatgc cgctctctat gagaggaagc tactgtctct ggaggtgcga aaacgtagac
     2461 gacggagtgg cagattgagg gcaatgaggc caaaataccc agtgattgcc caaccagctg
     2521 aaatgaatat taaaaccgag acagagagtg aagaggagga agaagttgga ttagacaatg
     2581 acgatgaaga gcaggaagca tcccaggagg agtctgcagg gtctcttgga gaaaatcaag
     2641 ccaagtacac accttcattg actgttatag tagaaaattc gcccagagac aatgcaatga
     2701 aagttgccga gtggactaat aaaggtgaac cgtgctgcaa aatcgaggct caggagccag
     2761 agtctaaatt caacctgatg cagattctcc aggacaatgg caatctaagc aaagtacagg
     2821 cccggttagc attctctgcc tatcttcagc atgtgcaaat ccgtctgaca aaagacagtg
     2881 gtggtcagac actcagtccc agctgggctg ccaaagagga tgaacagatg gaactggtgg
     2941 ttcgttttct caagcgagcg tcaagtaacc tccagcactc gctgaggatg gtattgccta
     3001 gtcggcggtt ggcacttctg gagcgcagga gaatccttgc ccaccagcta ggagacttta
     3061 tcggtgtgta caacaaggaa acagaacaaa tggctgaaaa gaaatcaaag aagaaacttg
     3121 aagaagagga agaagatggg gtgaacgcag aaagctttca ggagtttatt cgacaagcaa
     3181 gtgaggctga actggaggag gtgttgactt tttataccca aaaaaacaag tctgccagtg
     3241 tcttcctggg gactcactcg aaaagctcta agaacagcag tagttactct gatagtgggg
     3301 caaaaggtga tcaccctgag accatacaag aagtgaaaat aaagcaaccc aaacagcaac
     3361 aggcaacaga aatccacgcc gataaattat ctcgatttac cacttcatca ggaaaagagg
     3421 cgaaactagt gtataccaat tgctcttctt tctgcagtcc tgctgctgtt cttctgcaga
     3481 gactccccag ctcccatctg tcgtccgtga tcacaacctc tgccctctcg gcaggccctg
     3541 gccaccatgc ttctctatct caaatccctc cagctgtccc cagcctgcct caccagccag
     3601 cactgctact gagccccgtt cctgacaatg cccctccctc catacactct ggaacccaga
     3661 atgtaagccc tgcggggctg ccacgctgtc gatccggcag ctacaccatt ggcccctttt
     3721 cctctttcca aagtgccgca catatctata gccagaaact gtctcgtccc tcctcagcaa
     3781 aggcagcagg gtcatgtcat ccacacaagc atcattcagg aattgccaaa acacaaaaag
     3841 agggtgaaga tgtttcatta aacaggaggt acaaccaaag tctggttacg gctgaactcc
     3901 agcggctcgc ggagaagcag gcagccaggc agtattctcc agccagtcac atcagtctcc
     3961 tcactcagca ggtgacaaac ctgaatttgg ccagcagtgt cataaacaga agcagtgcat
     4021 ccaccccccc tacccttcgg cctgtcatca gtcctagtgg cccaacgtgg tctatacagc
     4081 cagacctcca cgcttctgag acccactcca gcccccctgg aagcaggagc ctccagacag
     4141 gtggctttgc ctgggagggc gaggtagaga acaacgcata cagcaagact acaggggtgg
     4201 tcccccagca caagtaccac cccacagcag gcagctacca gctccacttt gctctccagc
     4261 aactcgagca acaaaaactt cagtcccggc aacttctgga ccagagtcga gcccggcacc
     4321 aggcaatctt tggcagccag acactaccta actccagttt atggacaatg aataatggtc
     4381 caggctgcag aatttcaagt gccacaactg gtggccagaa gccaaacacc ctgccacaaa
     4441 aagtagtggc acctccaaat tcatccacct tggtctccaa gcctgcatcc aatcacaagc
     4501 aagtgctcag aaagccggca tcccagaggg cttccaaagg gtcctctgca gaagggcagc
     4561 tgaacggtct ccagagcagc cttaaccctg cagccttcat gcccatcacc aactccacag
     4621 gatctttgga agctccccaa gttattttcg caagatccaa acccttaccc acacagtctg
     4681 gagcccttgc tacagttata ggacagagaa aatcaaagtc ggtgaagtca gggacaattg
     4741 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4801 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4861 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4921 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4981 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5041 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5101 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5161 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5221 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5281 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5341 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5401 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     5461 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     5521 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5581 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5641 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5701 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5881 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     6121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     6181 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6241 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6301 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6361 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6421 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     6481 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     6541 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6601 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6661 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6721 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6781 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6841 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6901 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6961 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     7021 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     7081 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7141 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7201 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7261 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7321 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7381 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7441 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7501 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7561 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7621 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7681 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7741 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7801 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7861 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7921 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7981 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8041 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8101 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8161 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8221 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8281 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     8341 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8401 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8461 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8521 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8581 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8641 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8701 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8761 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8821 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8881 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8941 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     9001 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     9061 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9121 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9181 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9241 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9301 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9361 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9421 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9481 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9541 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9601 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9661 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9721 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9781 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9841 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9901 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9961 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
    10021 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
