LOCUS       V031624                 9950 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031624
VERSION     V031624
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..5440
                     /label="CPS1(NM_001122633)"
                     /note="CPS1(NM_001122633)"
                     /gene="CPS1"
     CDS             5447..5512
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    5550..5774
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5820..6248
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6262..6591
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6658..7452
                     /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    7626..7747
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7796..7812)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7820..7836
                     /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(7844..7874)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7889..7910
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8198..8783)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8954..9814)
                     /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(9815..9919)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgacgagga ttttgacagc
      961 tttcaaagtg gtgaggacac tgaagactgg ttttggcttt accaatgtga ctgcacacca
     1021 aaaatggaaa ttttcaagac ctggcatcag gctcctttct gtcaaggcac agacagcaca
     1081 cattgtcctg gaagatggaa ctaagatgaa aggttactcc tttggccatc catcctctgt
     1141 tgctggtgaa gtggttttta atactggcct gggagggtac ccagaagcta ttactgaccc
     1201 tgcctacaaa ggacagattc tcacaatggc caaccctatt attgggaatg gtggagctcc
     1261 tgatactact gctctggatg aactgggact tagcaaatat ttggagtcta atggaatcaa
     1321 ggtttcaggt ttgctggtgc tggattatag taaagactac aaccactggc tggctaccaa
     1381 gagtttaggg caatggctac aggaagaaaa ggttcctgca atttatggag tggacacaag
     1441 aatgctgact aaaataattc gggataaggg taccatgctt gggaagattg aatttgaagg
     1501 tcagcctgtg gattttgtgg atccaaataa acagaatttg attgctgagg tttcaaccaa
     1561 ggatgtcaaa gtgtacggca aaggaaaccc cacaaaagtg gtagctgtag actgtgggat
     1621 taaaaacaat gtaatccgcc tgctagtaaa gcgaggagct gaagtgcact tagttccctg
     1681 gaaccatgat ttcaccaaga tggagtatga tgggattttg atcgcgggag gaccggggaa
     1741 cccagctctt gcagaaccac taattcagaa tgtcagaaag attttggaga gtgatcgcaa
     1801 ggagccattg tttggaatca gtacaggaaa cttaataaca ggattggctg ctggtgccaa
     1861 aacctacaag atgtccatgg ccaacagagg gcagaatcag cctgttttga atatcacaaa
     1921 caaacaggct ttcattactg ctcagaatca tggctatgcc ttggacaaca ccctccctgc
     1981 tggctggaaa ccactttttg tgaatgtcaa cgatcaaaca aatgagggga ttatgcatga
     2041 gagcaaaccc ttcttcgctg tgcagttcca cccagaggtc accccggggc caatagacac
     2101 tgagtacctg tttgattcct ttttctcact gataaagaaa ggaaaagcta ccaccattac
     2161 atcagtctta ccgaagccag cactagttgc atctcgggtt gaggtttcca aagtccttat
     2221 tctaggatca ggaggtctgt ccattggtca ggctggagaa tttgattact caggatctca
     2281 agctgtaaaa gccatgaagg aagaaaatgt caaaactgtt ctgatgaacc caaacattgc
     2341 atcagtccag accaatgagg tgggcttaaa gcaagcggat actgtctact ttcttcccat
     2401 cacccctcag tttgtcacag aggtcatcaa ggcagaacag ccagatgggt taattctggg
     2461 catgggtggc cagacagctc tgaactgtgg agtggaacta ttcaagagag gtgtgctcaa
     2521 ggaatatggt gtgaaagtcc tgggaacttc agttgagtcc attatggcta cggaagacag
     2581 gcagctgttt tcagataaac taaatgagat caatgaaaag attgctccaa gttttgcagt
     2641 ggaatcgatt gaggatgcac tgaaggcagc agacaccatt ggctacccag tgatgatccg
     2701 ttccgcctat gcactgggtg ggttaggctc aggcatctgt cccaacagag agactttgat
     2761 ggacctcagc acaaaggcct ttgctatgac caaccaaatt ctggtggaga agtcagtgac
     2821 aggttggaaa gaaatagaat atgaagtggt tcgagatgct gatgacaatt gtgtcactgt
     2881 ctgtaacatg gaaaatgttg atgccatggg tgttcacaca ggtgactcag ttgttgtggc
     2941 tcctgcccag acactctcca atgccgagtt tcagatgttg agacgtactt caatcaatgt
     3001 tgttcgccac ttgggcattg tgggtgaatg caacattcag tttgcccttc atcctacctc
     3061 aatggaatac tgcatcattg aagtgaatgc cagactgtcc cgaagctctg ctctggcctc
     3121 aaaagccact ggctacccat tggcattcat tgctgcaaag attgccctag gaatcccact
     3181 tccagaaatt aagaacgtcg tatccgggaa gacatcagcc tgttttgaac ctagcctgga
     3241 ttacatggtc accaagattc cccgctggga tcttgaccgt tttcatggaa catctagccg
     3301 aattggtagc tctatgaaaa gtgtaggaga ggtcatggct attggtcgta cctttgagga
     3361 gagtttccag aaagctttac ggatgtgcca cccatctata gaaggtttca ctccccgtct
     3421 cccaatgaac aaagaatggc catctaattt agatcttaga aaagagttgt ctgaaccaag
     3481 cagcacgcgt atctatgcca ttgccaaggc cattgatgac aacatgtccc ttgatgagat
     3541 tgagaagctc acatacattg acaagtggtt tttgtataag atgcgtgata ttttaaacat
     3601 ggaaaagaca ctgaaaggcc tcaacagtga gtccatgaca gaagaaaccc tgaaaagggc
     3661 aaaggagatt gggttctcag ataagcagat ttcaaaatgc cttgggctca ctgaggccca
     3721 gacaagggag ctgaggttaa agaaaaacat ccacccttgg gttaaacaga ttgatacact
     3781 ggctgcagaa tacccatcag taacaaacta tctctatgtt acctacaatg gtcaggagca
     3841 tgatgtcaat tttgatgacc atggaatgat ggtgctaggc tgtggtccat atcacattgg
     3901 cagcagtgtg gaatttgatt ggtgtgctgt ctctagtatc cgcacactgc gtcaacttgg
     3961 caagaagacg gtggtggtga attgcaatcc tgagactgtg agcacagact ttgatgagtg
     4021 tgacaaactg tactttgaag agttgtcctt ggagagaatc ctagacatct accatcagga
     4081 ggcatgtggt ggctgcatca tatcagttgg aggccagatt ccaaacaacc tggcagttcc
     4141 tctatacaag aatggtgtca agatcatggg cacaagcccc ctgcagatcg acagggctga
     4201 ggatcgctcc atcttctcag ctgtcttgga tgagctgaag gtggctcagg caccttggaa
     4261 agctgttaat actttgaatg aagcactgga atttgcaaag tctgtggact acccctgctt
     4321 gttgaggcct tcctatgttt tgagtgggtc tgctatgaat gtggtattct ctgaggatga
     4381 gatgaaaaaa ttcctagaag aggcgactag agtttctcag gagcacccag tggtgctgac
     4441 aaaatttgtt gaaggggccc gagaagtaga aatggacgct gttggcaaag atggaagggt
     4501 tatctctcat gccatctctg aacatgttga agatgcaggt gtccactcgg gagatgccac
     4561 tctgatgctg cccacacaaa ccatcagcca aggggccatt gaaaaggtga aggatgctac
     4621 ccggaagatt gcaaaggctt ttgccatctc tggtccattc aacgtccaat ttcttgtcaa
     4681 aggaaatgat gtcttggtga ttgagtgtaa cttgagagct tctcgatcct tcccctttgt
     4741 ttccaagact cttggggttg acttcattga tgtggccacc aaggtgatga ttggagagaa
     4801 tgttgatgag aaacatcttc caacattgga ccatcccata attcctgctg actatgttgc
     4861 aattaaggct cccatgtttt cctggccccg gttgagggat gctgacccca ttctgagatg
     4921 tgagatggct tccactggag aggtggcttg ctttggtgaa ggtattcata cagccttcct
     4981 aaaggcaatg ctttccacag gatttaagat accccagaaa ggcatcctga taggcatcca
     5041 gcaatcattc cggccaagat tccttggtgt ggctgaacaa ttacacaatg aaggtttcaa
     5101 gctgtttgcc acggaagcca catcagactg gctcaacgcc aacaatgtcc ctgccacccc
     5161 agtggcatgg ccgtctcaag aaggacagaa tcccagcctc tcttccatca gaaaattgat
     5221 tagagatggc agcattgacc tagtgattaa ccttcccaac aacaacacta aatttgtcca
     5281 tgataattat gtgattcgga ggacagctgt tgatagtgga atccctctcc tcactaattt
     5341 tcaggtgacc aaactttttg ctgaagctgt gcagaaatct cgcaaggtgg actccaagag
     5401 tcttttccac tacaggcagt acagtgctgg aaaagcagca ctcgaggact acaaagacca
     5461 tgacggtgat tataaagatc atgacatcga ctacaaggat gacgatgaca agtagtgagg
     5521 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5581 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5641 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5701 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5761 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6061 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6121 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6181 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6241 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6421 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     6541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6601 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     6661 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6721 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6781 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6841 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6901 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6961 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7021 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7081 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7141 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7201 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7261 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7321 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7381 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7441 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7501 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7561 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7621 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7681 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7741 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     7801 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7861 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7921 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     8041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     8101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     8161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     8221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     8281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     8341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     8401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     8461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     8521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     8581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     8641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8701 cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta
     8761 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8821 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8881 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8941 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     9001 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     9061 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     9121 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     9181 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9241 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9301 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9361 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9421 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9481 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9541 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9601 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9661 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9721 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9781 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9841 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9901 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
