LOCUS       V021020                 8588 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021020
VERSION     V021020
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..3071
                     /label="DRD1(NM_000794)"
                     /note="DRD1(NM_000794)"
                     /gene="DRD1"
     promoter        3115..3326
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3339..3607
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3636..3645
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3666..4328
                     /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             4398..4451
                     /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             4452..5051
                     /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    5159..5280
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6567..6583)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6591..6607
                     /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(6615..6645)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6660..6681
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6969..7557)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7728..8588)
                     /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 accatgagga
     1741 ctctgaacac ctctgccatg gacgggactg ggctggtggt ggagagggac ttctctgttc
     1801 gtatcctcac tgcctgtttc ctgtcgctgc tcatcctgtc cacgctcctg gggaacacgc
     1861 tggtctgtgc tgccgttatc aggttccgac acctgcggtc caaggtgacc aacttctttg
     1921 tcatctcctt ggctgtgtca gatctcttgg tggccgtcct ggtcatgccc tggaaggcag
     1981 tggctgagat tgctggcttc tggccctttg ggtccttctg taacatctgg gtggcctttg
     2041 acatcatgtg ctccactgca tccatcctca acctctgtgt gatcagcgtg gacaggtatt
     2101 gggctatctc cagccctttc cggtatgaga gaaagatgac ccccaaggca gccttcatcc
     2161 tgatcagtgt ggcatggacc ttgtctgtac tcatctcctt catcccagtg cagctcagct
     2221 ggcacaaggc aaaacccaca agcccctctg atggaaatgc cacttccctg gctgagacca
     2281 tagacaactg tgactccagc ctcagcagga catatgccat ctcatcctct gtaataagct
     2341 tttacatccc tgtggccatc atgattgtca cctacaccag gatctacagg attgctcaga
     2401 aacaaatacg gcgcattgcg gccttggaga gggcagcagt ccacgccaag aattgccaga
     2461 ccaccacagg taatggaaag cctgtcgaat gttctcaacc ggaaagttct tttaagatgt
     2521 ccttcaaaag agaaactaaa gtcctgaaga ctctgtcggt gatcatgggt gtgtttgtgt
     2581 gctgttggct acctttcttc atcttgaact gcattttgcc cttctgtggg tctggggaga
     2641 cgcagccctt ctgcattgat tccaacacct ttgacgtgtt tgtgtggttt gggtgggcta
     2701 attcatcctt gaaccccatc atttatgcct ttaatgctga ttttcggaag gcattttcaa
     2761 ccctcttagg atgctacaga ctttgccctg cgacgaataa tgccatagag acggtgagta
     2821 tcaataacaa tggggccgcg atgttttcca gccatcatga gccacgaggc tccatctcca
     2881 aggagtgcaa tctggtttac ctgatcccac atgctgtggg ctcctctgag gacctgaaaa
     2941 aggaggaggc agctggcatc gccagaccct tggagaagct gtccccagcc ctatcagtca
     3001 tattggacta tgacactgac gtctctctgg agaagatcca acccatcaca caaaacggtc
     3061 agcacccaac ctgagcggcc gcaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     3121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     3181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     3241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     3301 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     3361 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     3421 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     3481 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     3541 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     3601 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg
     3661 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg
     3721 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga
     3781 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct
     3841 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca
     3901 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga
     3961 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg
     4021 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg
     4081 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc
     4141 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca
     4201 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc
     4261 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg
     4321 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg
     4381 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga
     4441 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4501 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4561 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4621 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4681 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4741 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4801 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4861 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4921 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4981 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5041 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     5101 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcagttaa
     5161 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5221 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5281 tcatgtctgg aattgactca aatgatgtca attagtctat cagaagctat ctggtctccc
     5341 ttccggggga caagacatcc ctgtttaata tttaaacagc agtgttccca aactgggttc
     5401 ttatatccct tgctctggtc aaccaggttg cagggtttcc tgtcctcaca ggaacgaagt
     5461 ccctaaagaa acagtggcag ccaggtttag ccccggaatt gactggattc cttttttagg
     5521 gcccattggt atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
     5581 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc
     5641 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc
     5701 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc
     5761 tgtccctgat atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     5821 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     5881 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     5941 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     6001 attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca
     6061 gactatcttt ctagggttaa tctagctgca tcaggatcat atcgtcgggt cttttttccg
     6121 gctcagtcat cgcccaagct ggcgctatct gggcatcggg gaggaagaag cccgtgcctt
     6181 ttcccgcgag gttgaagcgg catggaaaga gtttgccgag gatgactgct gctgcattga
     6241 cgttgagcga aaacgcacgt ttaccatgat gattcgggaa ggtgtggcca tgcacgcctt
     6301 taacggtgaa ctgttcgttc aggccacctg ggataccagt tcgtcgcggc ttttccggac
     6361 acagttccgg atggtcagcc cgaagcgcat cagcaacccg aacaataccg gcgacagccg
     6421 gaactgccgt gccggtgtgc agattaatga cagcggtgcg gcgctgggat attacgtcag
     6481 cgaggacggg tatcctggct ggatgccgca gaaatggaca tggatacccc gtgagttacc
     6541 cggcgggcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6601 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6661 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6721 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6781 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6841 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6901 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6961 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7021 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7081 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7141 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7201 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7261 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7321 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7381 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7441 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7501 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7561 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7621 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7681 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7741 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7801 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7861 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7921 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7981 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8041 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8101 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8161 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8221 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8281 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8341 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8401 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8461 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8521 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8581 atactcat
//
