LOCUS       V041616                 6370 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041616
VERSION     V041616
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             928..1806
                     /label="NUS1(NM_138459)"
                     /note="NUS1(NM_138459)"
                     /gene="NUS1"
     CDS             1813..1836
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             1843..1932
                     /codon_start=1
                     /product="two Strep-Tag®II moieties connected by a linker
                     for enhanced binding to Strep-Tactin®,&nbsp;an engineered
                     form of streptavidin (Schmidt et al., 2013)"
                     /transl_table=1
                     /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK"
                     /label="Twin-Strep-tag"
     polyA_signal    1970..2194
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2240..2668
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2682..3011
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3078..3872
                     /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    4046..4167
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4216..4232)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4240..4256
                     /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(4264..4294)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4309..4330
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4618..5203)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5374..6234)
                     /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(6235..6339)
                     /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 aagctgaatt cgccaccatg acggggctgt acgagctggt gtggcgggtg
      961 ctgcacgcgc tgctctgtct gcaccgcacg ctcacctcct ggctccgcgt tcggttcggc
     1021 acctggaact ggatctggcg gcgctgctgc cgcgccgcct ctgccgcggt cctagcgccg
     1081 ctcggcttca cgctccgcaa gcccccggca gtcggcagga accgccgtca ccaccggcac
     1141 ccgcgcgggg ggtcgtgcct ggcagccgca caccaccgga tgcgctggcg cgcggacggt
     1201 cgttccttgg agaagctgcc tgtgcatatg ggcctggtga tcaccgaggt ggagcaggaa
     1261 cccagcttct cggacatcgc gagcctcgtg gtgtggtgta tggccgtggg catctcctac
     1321 attagcgtct acgaccacca aggtattttc aaaagaaata attccagatt gatggatgaa
     1381 attttaaaac aacagcaaga acttctgggc ctagattgtt caaaatactc accagaattt
     1441 gcaaatagta atgacaaaga tgatcaagtt ttaaattgcc atttggcagt gaaggtgctg
     1501 tctccggaag atggaaaagc agatattgta agagctgctc aggacttttg ccagttagta
     1561 gcccagaagc aaaagagacc cacagatttg gatgtagata cgttagccag tttacttagt
     1621 tcaaatggtt gtcctgatcc tgatttagta ttgaagttcg gtcctgtgga cagcacatta
     1681 ggctttcttc cctggcacat cagattgact gagattgtct ctttgccttc ccacctaaac
     1741 atcagttatg aggacttttt ctctgccctt cgtcaatatg cagcctgtga acagcgtctg
     1801 ggaaagctcg aggattacaa ggatgacgac gataagggat ccagcgcctg gtcccaccct
     1861 cagtttgaga agggcggagg ctctggcggc ggaagcggag gatctgcttg gagccacccc
     1921 cagttcgaaa agtgatgagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     1981 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2041 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2101 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2161 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2221 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2281 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2341 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2401 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2461 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2521 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2581 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2641 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2761 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2821 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2881 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2941 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3001 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3061 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3121 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3181 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3241 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3301 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3361 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3421 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3481 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3541 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3601 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3661 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3721 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3781 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3841 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3901 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3961 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4021 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4141 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     5161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6361 acctgacgtc
//
