LOCUS       V041809                 8409 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041809
VERSION     V041809
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..3089
                     /label="Fam193b(NM_145382)"
                     /note="Fam193b(NM_145382)"
                     /gene="Fam193b"
     CDS             3096..3161
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3185..3773
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3839..3887
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4089..4517
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4531..4860
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4927..5721
                     /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    5897..6018
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6067..6083)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6091..6107
                     /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(6115..6145)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6160..6181
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6469..7057)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7228..8088)
                     /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(8089..8193)
                     /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 aaagctcgtc aagaatctcc
      781 taggcgagat gcctctgtgg gtctgccaga gctgccgaaa gagcatggag gaggaggaga
      841 ggcagacagg tggagaccat gcagtggcga tctccttgtc acacacatcc tgcaaatcac
      901 agtcttgtgg gggtgactct cattcatcct cgtcctcttc atcatcctca tcttcctcct
      961 cctcttcctg ccatgggaac tcaggggact gggatcccag ctcgttcctg tcagcacata
     1021 agctctcagg cctctggaac tccccacact ccagcggggc tgtgccaggc agctcgcttg
     1081 ggagtcctcc tgccatcctt ggtgaggctt tccctgtctc ggagcaccac cagcactcag
     1141 accttactgc tccccctaac agccccactg gtcctccccc ccacccagca tctctgattc
     1201 cttctcaccc tggttccttc agctcggcgt cctacccacc cccgctgccc accaccccag
     1261 tggcaccttt ccccgctcag gcttcagagt gccctatggc tgctgccact gccacccacc
     1321 cctcggggcc atgtcagagc cctcatccgc cctccaccag catgccactc ctgaagatgc
     1381 ctccaccact ctcggggtgc agccacccct gcagcgggca ctgcagcggg cactgtggtg
     1441 gtcctcttct cccaccaccg agctctcagc cgctccctag cactcacagc agggaccctg
     1501 ggtgcaaggg gcacaaattt gcacacagtg gcctggcctg tgaggcagat gagggtctgg
     1561 gcgaggaaga ggatagcagc tctgagcgca gctcttgcac ctcgtcctcc acccaccccc
     1621 gcgatgggaa gttctgtgac tgctgctact gtgagttctt cggccacaat gcgccactcg
     1681 ctgccccgac gagccggaat tatactgaga tccgggagaa gctccgctca aggctgacca
     1741 ggcggaaaga ggagctgccc atgaaggggg gcaccctggg cgggatccct ggggagcctg
     1801 cggtggacca ccgagatgtg gatgagctgc tggaattcat caacagcacg gagcccaaag
     1861 tccccaacag cgccagggct gccaagcggg cccggcacaa gctgaaaaag aaggaaaaag
     1921 agaaggctcg gttggccaca gaagctctga agcaggtgaa tcgtgtttct ggaagccagg
     1981 agccaaggcc tgccagggag aggctcctag agtggcctga ccaggaactg gaccgcgtca
     2041 atagcttctt gagcagccgc ttgcaggaga tcaagagcac tgtcaaggac tccctctgtg
     2101 ccagcctgag tatgtgtgag ctcagcgtgg agagcagtgg gtttaaggag gggactgtgg
     2161 aagctcagac cctcactcct tcagacctca gtggctcctc tcagaaacgg cctgacatca
     2221 accttgacct gtcccctttg accttgggtt cccctcagag ccacacgtta caagccccaa
     2281 gtgaaccagt cccaccttgg gcagaaagga gagaccctca cccaccacca tggacagagg
     2341 tgaggggccc acctcctggt atccctgaga atgggcttgt gagaagactc aacactgtgc
     2401 ccaacctgtc cagggtgatc tgggtcaaga cacccaagcc aggcaaccca agctctgagg
     2461 aaagttcaaa ggaagttccc agttgcaaac aggaattatc tgagcctgtg gccacaggtg
     2521 gaaagccaaa aaagagcaag agacagggaa gtcaagcaaa gaagactttg gcaagtccag
     2581 ccccctggtc accagccaat ctagaggctt ccggtgccaa gagccaggta tctagcccca
     2641 agcagcccag caagggctca gagcctgcca aagtgggcag tggtgcagaa cctggagagg
     2701 gcagtccggg gagccggcca ggaccaattc aggctgacag ccccaaaact gacaagaagg
     2761 gtagttcctg gcagaattgg ccaggtgggg ccaaggcacg gactctggag caggaatctg
     2821 agcagacccc tggtccagca aggccacaga gcttgtccca aggcaagggc cgcagccgcc
     2881 ggagccgaaa caagcaggag aagtcagcct cctccttgga cgatgtgttc ctgcccaaag
     2941 acttggatgg ggtggagatg gatgagacag accgagaggt ggagtacttc aagaggttct
     3001 gtttggattc tgcaaagcaa actcgtcaga aagttgctgt gaactggacc aacttcagcc
     3061 tcaagaaaac cactcctagc acagctcagg gatccgacta caaagaccat gacggtgatt
     3121 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct
     3181 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3241 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3301 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3361 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3421 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3481 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3541 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3601 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3661 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3721 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3781 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3841 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     3901 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3961 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4021 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4081 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4141 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     4261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4381 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4441 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4501 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4561 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4621 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4681 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4741 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4801 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4861 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4921 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4981 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5041 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5101 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5161 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5221 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5281 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5341 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5401 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5461 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5521 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5581 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5641 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5701 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     5761 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5821 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5881 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5941 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6001 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6061 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6121 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6181 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6241 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6301 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6361 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6421 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6481 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6541 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6601 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6661 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6721 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6781 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6841 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6901 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6961 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7021 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7081 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7141 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7201 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7261 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7321 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7381 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7441 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7501 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7561 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7621 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7681 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7741 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7801 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7861 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7921 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7981 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8041 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8101 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8161 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8221 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     8281 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     8341 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8401 ctgcttagg
//
