LOCUS       V044325                 9359 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044325
VERSION     V044325
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(1..105)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     728..744
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        1442..1645
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             1734..2156
                     /label="1600014C10Rik(NM_001085385)"
                     /note="1600014C10Rik(NM_001085385)"
                     /gene="1600014C10Rik"
     CDS             2193..3845
                     /EC_number=""
                     /codon_start=1
                     /gene="luc+"
                     /note="enhanced&nbsp;luc+ version of the luciferase gene"
                     /product="firefly luciferase"
                     /protein_id=""
                     /transl_table=1
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV*"
                     /label="luciferase"
     promoter        3886..4097
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4110..4378
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4407..4416
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4437..5099
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             5169..5222
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             5223..5822
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5930..6051
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7338..7354)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7362..7378
                     /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(7386..7416)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7431..7452
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7740..8328)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8499..9359)
                     /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"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1501 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1561 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1621 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1681 tccacgctgt tttgacctcc atagaagatt ctagagctag cgaattcgcc accatgccca
     1741 tcatggtaga tgacatcatg aggcttctgt gctccatttc ccaggagagg aagatgaagg
     1801 cggccgtcaa gcactctgga aagggtgcca tggttgcggg ggcaatggcc ttcgtcggtg
     1861 gcttggttgg tggcccacca ggaatagctg ttggggggac cgttgggggc ctgttaggtg
     1921 cctggatgac gagtggacag ttcaagcccg ttcctcagat cttaatggag ctgccccccg
     1981 ctgaacagcg gaaacttgtt aacgaagcca tggccatcat cgggaacttg gactggacag
     2041 acgcagtgca gctgactgct ctggtcatga gcaaccaggc catgcagcag aggctcttgg
     2101 ccatgctaac gacctatgtc accaaggagc tgcaggctga gatccgctat gaggacggtg
     2161 gaggcggatc aggaggcggt ggatccgcca ccatggaaga cgccaaaaac ataaagaaag
     2221 gcccggcgcc attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta
     2281 tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg
     2341 acatcactta cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat
     2401 atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta
     2461 tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata
     2521 atgaacgtga attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca
     2581 aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta
     2641 ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat
     2701 ctcatctacc tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca
     2761 agacaattgc actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc
     2821 tgcctcatag aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc
     2881 aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt
     2941 ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag
     3001 aagagctgtt tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa
     3061 ccctattctc cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac
     3121 acgaaattgc ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga
     3181 ggttccatct gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc
     3241 tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg
     3301 aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac
     3361 tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg
     3421 ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg
     3481 aacacttctt catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg
     3541 ctcccgctga attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg
     3601 caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg
     3661 gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga
     3721 aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac
     3781 tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg
     3841 tgtaagcggc cgcaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     3901 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     3961 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4021 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4081 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     4141 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     4201 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     4261 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     4321 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     4381 agctgtgacc ggcgcctacg ctagacgcca ccatggagag cgacgagagc ggcctgcccg
     4441 ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc gagctggtgg
     4501 gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag agcaccaaag
     4561 gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc ttctaccact
     4621 tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac aacggcggct
     4681 acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg agcttcagct
     4741 accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc ggcttccccg
     4801 aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg gagcacctgc
     4861 accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc ctgcgcgacg
     4921 gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc atccacccca
     4981 gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag ctgcacagca
     5041 acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc gccttcgcca
     5101 gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc ggctccaccg
     5161 gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     5221 ctatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     5281 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     5341 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     5401 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     5461 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     5521 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     5581 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     5641 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     5701 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     5761 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     5821 gaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5881 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcagtta acttgtttat
     5941 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6001 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6061 gaattgactc aaatgatgtc aattagtcta tcagaagcta tctggtctcc cttccggggg
     6121 acaagacatc cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc
     6181 ttgctctggt caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga
     6241 aacagtggca gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg
     6301 tatggctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt
     6361 tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg
     6421 gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc
     6481 ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga
     6541 tatctataac aagaaaatat atatataata agttatcacg taagtagaac atgaaataac
     6601 aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt
     6661 ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc
     6721 ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct
     6781 atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt
     6841 tctagggtta atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca
     6901 tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga
     6961 ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg
     7021 aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga
     7081 actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg
     7141 gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg
     7201 tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg
     7261 gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg
     7321 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7381 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7441 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7501 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7561 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7621 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7681 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7741 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7801 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7861 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7921 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7981 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8041 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8101 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8161 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8221 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8281 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8341 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8401 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8461 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8521 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8581 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8641 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8701 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8761 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8821 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8881 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8941 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     9001 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     9061 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     9121 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     9181 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9241 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9301 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcat
//
