LOCUS       V042347                 8004 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042347
VERSION     V042347
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..2684
                     /label="ASNSD1(NM_019048)"
                     /note="ASNSD1(NM_019048)"
                     /gene="ASNSD1"
     CDS             2691..2756
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2780..3368
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3434..3482
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3684..4112
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4126..4455
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4522..5316
                     /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    5492..5613
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5662..5678)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5686..5702
                     /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(5710..5740)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5755..5776
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6064..6652)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6823..7683)
                     /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(7684..7788)
                     /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 ccaccatgtg tggcatttgt tgttctgtaa
      781 acttttctgc tgagcatttc agtcaagatt taaaagagga cttactatat aatcttaaac
      841 agcggggacc caatagtagt aaacaattgt taaagtctga tgttaactac cagtgtttat
      901 tttctgctca cgtcctacac ttgaggggtg ttttgactac ccagcctgtg gaagatgaaa
      961 gaggcaatgt gtttctatgg aatggagaaa tttttagtgg aataaaggtt gaagctgaag
     1021 agaatgacac tcaaattttg tttaattatc tttcctcctg taagaatgaa tctgagattt
     1081 tgtcactctt ctcagaagta caaggtccct ggtcatttat atattatcaa gcatctagtc
     1141 attatttatg gtttggtagg gatttttttg gtcgccgtag cttgctttgg cattttagta
     1201 atttgggcaa gagtttctgc ctctcttcag ttggcaccca aacatctgga ttggcaaatc
     1261 agtggcaaga agttccagca tctggacttt tcagaattga tcttaagtct actgtcattt
     1321 ccggatgcat tattttacaa ctgtatcctt ggaaatatat ttctagggag aatattattg
     1381 aagaaaatgt taatagcctg agtcaaattt cagcagactt accagcattt gtatcagtgg
     1441 tagcaaatga agccaaactg tatcttgaaa aacctgttgt tcctttaaat atgatgttgc
     1501 cacaagctgc attggagact cattgcagta atatttccaa tgtgccacct acaagagaga
     1561 tacttcaagt ctttcttact gatgtacaca tgaaggaagt aattcagcag ttcattgatg
     1621 tcctgagtgt agcagtcaag aaacgtgtct tgtgtttacc tagggatgaa aacctgacag
     1681 caaatgaagt tttgaaaacg tgtgatagga aagcaaatgt tgcaatcctg ttttctgggg
     1741 gcattgattc catggttatt gcaacccttg ctgaccgtca tattccttta gatgaaccaa
     1801 ttgatcttct taatgtagct ttcatagctg aagaaaagac catgccaact acctttaaca
     1861 gagaagggaa taaacagaaa aataaatgtg aaataccttc agaagaattc tctaaagatg
     1921 ttgctgctgc tgctgctgac agtcctaata aacatgtcag tgtaccagat cgaatcacag
     1981 gaagggcggg actaaaggaa ctacaagctg ttagcccttc ccgaatttgg aattttgttg
     2041 aaattaatgt ttctatggaa gaactgcaga aattaagaag aactcgaata tgtcacttaa
     2101 ttcggccatt ggatacagtt ttggatgata gcattggctg tgcagtctgg tttgcttcta
     2161 gaggaattgg ttggttagtg gcccaggaag gagtgaaatc ctatcagagc aatgcaaagg
     2221 tagttctcac tggaattggt gcagatgagc aacttgcagg ttattctcgt catcgtgtcc
     2281 gctttcagtc gcatgggctg gaaggattga ataaggaaat aatgatggaa ctgggtcgaa
     2341 tttcttctag aaatcttggt cgtgatgaca gagttattgg tgatcatgga aaagaagcaa
     2401 gatttccttt cctggatgaa aatgttgtct cctttctaaa ttctctgccg atttgggaaa
     2461 aagcaaactt gactttaccc cgaggaattg gtgaaaaatt acttttacgc cttgcagctg
     2521 tggaacttgg tcttacagcc tctgctcttc tgcccaaacg ggccatgcag tttggatcaa
     2581 gaattgcaaa aatggaaaaa attaatgaaa aggcatctga taaatgtgga cggctccaaa
     2641 tcatgtcctt agaaaatctt tctattgaaa aggagactaa attgggatcc gactacaaag
     2701 accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt
     2761 gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2821 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2881 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2941 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3001 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3061 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3121 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3181 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3241 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3301 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3361 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3421 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3481 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3541 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3601 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3661 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3721 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3781 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3841 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3901 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3961 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4021 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4081 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4201 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4321 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4381 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4441 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4501 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4561 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4621 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4681 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4741 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4801 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4861 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4921 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4981 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5041 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5101 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5161 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5221 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5281 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5341 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5401 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5461 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5521 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5581 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5641 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5701 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5761 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5821 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5881 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5941 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6001 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6241 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6481 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7801 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7861 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7921 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     7981 aattgcatga agaatctgct tagg
//
