LOCUS       V030449                 8778 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030449
VERSION     V030449
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..3458
                     /label="SLFN11(NM_001104587)"
                     /note="SLFN11(NM_001104587)"
                     /gene="SLFN11"
     CDS             3465..3530
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3554..4142
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4208..4256
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4458..4886
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4900..5229
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5296..6090
                     /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    6266..6387
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6436..6452)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6460..6476
                     /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(6484..6514)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6529..6550
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6838..7426)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7597..8457)
                     /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(8458..8562)
                     /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 ccaccatgga ggcaaatcag tgccccctgg
      781 ttgtggaacc atcttaccca gacctggtca tcaatgtagg agaagtgact cttggagaag
      841 aaaacagaaa aaagctgcag aaaattcaga gagaccaaga gaaggagaga gttatgcggg
      901 ctgcatgtgc tttattaaac tcaggaggag gagtgattcg aatggccaag aaggttgagc
      961 atcccgtgga gatgggactg gatttagaac agtctttgag agagcttatt cagtcttcag
     1021 atctgcaggc tttctttgag accaagcaac aaggaaggtg tttttacatt tttgttaaat
     1081 cttggagcag tggccctttc cctgaagatc gctctgtcaa gccccgcctt tgcagcctca
     1141 gttcttcatt ataccgtaga tctgagacct ctgtgcgttc catggactca agagaggcat
     1201 tctgtttcct gaagaccaaa aggaagccaa aaatcttgga agaaggacct tttcacaaaa
     1261 ttcacaaggg tgtataccaa gagctcccta actcggatcc tgctgaccca aactcggatc
     1321 ctgctgacct aattttccaa aaagactatc ttgaatatgg tgaaatcctg ccttttcctg
     1381 agtctcagtt agtagagttt aaacagttct ctacaaaaca cttccaagaa tatgtaaaaa
     1441 ggacaattcc agaatacgtc cctgcatttg caaacactgg aggaggctat ctttttattg
     1501 gagtggatga taagagtagg gaagtcctgg gatgtgcaaa agaaaatgtt gaccctgact
     1561 ctttgagaag gaaaatagaa caagccatat acaaactacc ttgtgttcat ttttgccaac
     1621 cccaacgccc gataaccttc acactcaaaa ttgtgaatgt gttaaaaagg ggagagctct
     1681 atggctatgc ttgcatgatc agagtaaatc ccttctgctg tgcagtgttc tcagaagctc
     1741 ccaattcatg gatagtggag gacaagtacg tctgcagcct gacaaccgag aaatgggtag
     1801 gcatgatgac agacacagat ccagatcttc tacagttgtc tgaagatttt gaatgtcagc
     1861 tgagtctatc tagtgggcct ccccttagca gaccagtgta ctccaagaaa ggcctggaac
     1921 ataaaaagga actccagcaa cttttatttt cagtcccacc aggatatttg cgatatactc
     1981 cagagtcact ctggagggac ctgatctcag agcacagagg actagaggag ttaataaata
     2041 agcaaatgca acctttcttt cggggaattt tgatcttctc tagaagttgg gctgtggacc
     2101 tgaacttgca ggagaagcca ggagtcatct gtgatgctct gctgatagca cagaacagca
     2161 cccccattct ctacaccatt ctcagggagc aagatgcaga gggccaggac tactgcactc
     2221 gcaccgcctt tactttgaag cagaagctag tgaacatggg gggctacacc gggaaggtgt
     2281 gtgtcagggc caaggtcctc tgcctgagtc ctgagagcag cgcagaggcc ttggaggctg
     2341 cagtgtctcc gatggattac cctgcgtcct atagccttgc aggcacccag cacatggaag
     2401 ccctgctgca gtccctcgtg attgtcttac tcggcttcag gtctctcttg agtgaccagc
     2461 tcggctgtga ggttttaaat ctgctcacag cccagcagta tgagatattc tccagaagcc
     2521 tccgcaagaa cagagagttg tttgtccacg gcttacctgg ctcagggaag accatcatgg
     2581 ccatgaagat catggagaag atcaggaatg tgtttcactg tgaggcacac agaattctct
     2641 acgtttgtga aaaccagcct ctgaggaact ttatcagtga tagaaatatc tgccgagcag
     2701 agacccggaa aactttccta agagaaaact ttgaacacat tcaacacatc gtcattgacg
     2761 aagctcagaa tttccgtact gaagatgggg actggtatgg gaaggcaaaa agcatcactc
     2821 ggagagcaaa gggtggccca ggaattctct ggatctttct ggattacttt cagaccagcc
     2881 acttggattg cagtggcctc cctcctctct cagaccaata tccaagagaa gagctcacca
     2941 gaatagttcg caatgcagat ccaatagcca agtacttaca aaaagaaatg caagtaatta
     3001 gaagtaatcc ttcatttaac atccccactg ggtgcctcga ggtatttcct gaagccgaat
     3061 ggtcccaggg tgttcaggga accttacgaa ttaagaaata cttgactgtg gagcaaataa
     3121 tgacctgtgt ggcagacacg tgcaggcgct tctttgatag gggctattct ccaaaggatg
     3181 ttgctgtgct tgtcagcacc gcaaaagaag tggagcacta taagtatgag ctcttgaaag
     3241 caatgaggaa gaaaagggtg gtgcagctca gtgatgcatg tgatatgttg ggtgatcaca
     3301 ttgtgttgga cagtgttcgg cgattctcag gcctggaaag gagcatagtg tttgggatcc
     3361 atccaaggac agctgaccca gctatcttac ccaatgttct gatctgtctg gcttccaggg
     3421 caaaacaaca cctgtatatt tttccgtggg gtggccatgg atccgactac aaagaccatg
     3481 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     3541 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3601 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3661 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3721 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3781 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3841 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3901 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3961 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4021 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4081 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4141 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4201 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     4261 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4321 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4381 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     4441 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4801 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     4861 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4981 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5041 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5101 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5161 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5221 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     5281 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5341 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5401 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5461 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5521 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5581 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5641 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5701 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5761 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5821 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5881 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5941 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6001 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6061 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6121 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     6181 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     6241 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     6301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6361 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     6421 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     6481 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     6541 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     6601 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6661 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7021 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7261 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8581 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     8641 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8701 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8761 atgaagaatc tgcttagg
//
