LOCUS       V019304                 7076 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019304
VERSION     V019304
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..2572
                     /label="Nap1l3(NM_138742)"
                     /note="Nap1l3(NM_138742)"
                     /gene="Nap1l3"
     CDS             2573..2638
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2676..2900
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2946..3374
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3388..3717
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3784..4578
                     /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    4752..4873
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4922..4938)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4946..4962
                     /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(4970..5000)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5015..5036
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5324..5909)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6080..6940)
                     /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(6941..7045)
                     /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 atggcagaag cggatcctaa
      961 aatggtcaca gaacctggtg cccatggggt tgctgaagag gcgatggcta gcacagcttg
     1021 tgattctggg gatgaatctg acagcaatag ctctagcagt accaatagtt gcagcagcag
     1081 cggcagcagc agcagcggca gcagcagtag cagcagcagc agtagcagca gcagcagcag
     1141 cagcagcagt agcagtagcg gtagcagtgg cagcagcagc aatggcagtc atttgaaccg
     1201 aaagaagagg gtacctgagc cttccagaag ggcccagcga cgtccctccg ggaaactttt
     1261 cttggataag ctgccccaag ccgtaagaaa tcgggtgcag gcactcagaa atattcagaa
     1321 tgagtgtgac aaggtagaca ccttgttctt aagggcaatt catgatcttg aaagaaagta
     1381 tgctgaactc aataagcctc tatatgataa gcgttttcag atcataaatg cagaatatga
     1441 gcctacagag gaagaatgtg aatggaattc agaagaagag ttcagtggtg atgaagaaat
     1501 gcaggatgac acacctaatg aaatgccacc cttagagggt gaggaggaag aagaaagctg
     1561 taatgaaaaa gctgaagtga aggaagaagg aacacatgtt ccagaagaag ttcctgaggc
     1621 aaaagttgaa gaagaggagg ctcccaaaga aactcctgag gtgaaaactg aagaaaaaga
     1681 cattccaaaa gaaggtgctg aagaaaaagc tgaagaacag gaatcctcta aagaaattcc
     1741 tgaggtaaaa ggtgaagaaa aagcagactc tacggattgt atagatatag ctcctgaaga
     1801 aaaagaagac gtcaaagaag ttacccaggc aaatacagaa aataaggatc aacctacaga
     1861 agaatttaca ccaagggctc cagcaagaga ggctcaaaaa agggtccctg agacaaggcc
     1921 tgaagaagga gtcaatatta aaagggctcg aaagggaaaa cctaagaaag aagatcctaa
     1981 aggtattcct gactactggc tgactgtttt aaagaatgtt gataagcttg ggcctatgat
     2041 tcaaaagtgt gatgaaccca ttttgaagtt cttatctgat gtgagcctga agttctcaaa
     2101 ccctggccag cctattggtt acacttttga atttcatttc ctacctaacc catacttcag
     2161 aaatgagctc ctgatgaaga catacataat aaggtcaaaa ccagatcact acgacccgtt
     2221 cttcgcgtgg ggatgggaaa ttgaagagtg taaaggctgc aaaatagact ggagacgagg
     2281 aaaagatgtt acggtgacaa ccacccggag tcgccctggt attacagggg aaattgaagt
     2341 ccagccaaga gtggttccta atgcatcctt cttcaatttc ttcagtcctc ctgagattcc
     2401 tttgattggg aagctggaac caagagaaga tgctatcctt gatgaggact ttgagattgg
     2461 tcaaattttg catgataatg tcatcttgaa gtcaatctat tacttcacag gagaaatcaa
     2521 tgatccctac taccatgact tcagggatta tggaaatagg aagtactaca aggactacaa
     2581 agaccatgac ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta
     2641 gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2701 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2761 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2821 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2881 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2941 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3001 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3241 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3301 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3361 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3601 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3661 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3721 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3781 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3841 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3901 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3961 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4021 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4081 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4141 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4201 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4261 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4321 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4381 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4441 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4501 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4561 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4621 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4681 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4741 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4861 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4981 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5761 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc
     5881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5941 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
