LOCUS       V041242                 7800 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041242
VERSION     V041242
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..2276
                     /label="Atf3(NM_007498)"
                     /note="Atf3(NM_007498)"
                     /gene="Atf3"
     promoter        2327..2538
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             2551..2819
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      2848..2857
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2878..3540
                     /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             3610..3663
                     /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             3664..4263
                     /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    4371..4492
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5779..5795)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5803..5819
                     /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(5827..5857)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5872..5893
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6181..6769)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6940..7800)
                     /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 accatgatgc
     1741 ttcaacatcc aggccaggtc tctgcctcag aagtcagtgc gaccgccatt gtcccctgcc
     1801 tctcacctcc tgggtcactg gtatttgagg attttgctaa cctgacaccc tttgtcaagg
     1861 aagagctgag attcgccatc cagaataaac acctctgcca tcggatgtcc tctgcgctgg
     1921 agtcagttac cgtcaacaac agacccctgg agatgtcagt caccaagtct gaggcggccc
     1981 ctgaagaaga tgagaggaaa aggaggcggc gagaaagaaa taaaattgct gctgccaagt
     2041 gtcgaaacaa gaaaaaggag aagacagagt gcctgcagaa agagtcagag aaactggaga
     2101 gtgtgaatgc tgagctgaag gcccagattg aggagctgaa gaatgagaaa cagcatttga
     2161 tatacatgct caacctgcac cggcccacct gcatcgtccg ggctcagaat ggacggacac
     2221 cggaagacga gaggaacctc tttatccaac agataaaaga aggaacattg cagagctaag
     2281 gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     2341 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     2401 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2461 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2521 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     2581 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     2641 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     2701 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     2761 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     2821 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc
     2881 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     2941 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa
     3001 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     3061 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     3121 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     3181 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     3241 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     3301 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     3361 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     3421 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     3481 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     3541 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc
     3601 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     3661 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     3721 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     3781 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     3841 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     3901 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     3961 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4021 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4081 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4141 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4201 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     4261 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4321 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcagtt aacttgttta
     4381 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4441 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4501 ggaattgact caaatgatgt caattagtct atcagaagct atctggtctc ccttccgggg
     4561 gacaagacat ccctgtttaa tatttaaaca gcagtgttcc caaactgggt tcttatatcc
     4621 cttgctctgg tcaaccaggt tgcagggttt cctgtcctca caggaacgaa gtccctaaag
     4681 aaacagtggc agccaggttt agccccggaa ttgactggat tcctttttta gggcccattg
     4741 gtatggcttt ttccccgtat ccccccaggt gtctgcaggc tcaaagagca gcgagaagcg
     4801 ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg ggctgtcccc gcacgctgcc
     4861 ggctcgggga tgcgggggga gcgccggacc ggagcggagc cccgggcggc tcgctgctgc
     4921 cccctagcgg gggagggacg taattacatc cctgggggct ttgggggggg gctgtccctg
     4981 atatctataa caagaaaata tatatataat aagttatcac gtaagtagaa catgaaataa
     5041 caatataatt atcgtatgag ttaaatctta aaagtcacgt aaaagataat catgcgtcat
     5101 tttgactcac gcggtcgtta tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg
     5161 cctcacggga gctccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc
     5221 tatttagaaa gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct
     5281 ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc
     5341 atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg
     5401 aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc
     5461 gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg
     5521 aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc
     5581 ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc
     5641 gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg
     5701 ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc
     5761 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5821 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5881 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5941 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6601 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6841 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6901 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6961 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7021 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7081 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7141 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7201 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7261 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7321 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7381 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7441 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7501 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7561 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7621 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7681 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7741 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
//
