LOCUS       V032181                 7836 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032181
VERSION     V032181
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..3128
                     /label="TACR3(NM_001059)"
                     /note="TACR3(NM_001059)"
                     /gene="TACR3"
     promoter        3173..3384
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3397..3665
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3700..4299
                     /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    4407..4528
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5815..5831)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5839..5855
                     /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(5863..5893)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5908..5929
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6217..6805)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6976..7836)
                     /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 accatggcca
     1741 ctctcccagc agcagaaacc tggatagacg ggggtggagg cgtgggtgca gacgccgtga
     1801 acctgaccgc ctcgctagct gccggggcgg ccacgggggc agttgagact gggtggctgc
     1861 aactgctgga ccaagctggc aacctctcct cctccccttc cgcgctggga ctgcctgtgg
     1921 cttcccccgc gccctcccag ccctgggcca acctcaccaa ccagttcgtg cagccgtcct
     1981 ggcgcatcgc gctctggtcc ctggcgtatg gtgtggtggt ggcagtggca gttttgggaa
     2041 atctcatcgt catctggatc atcctggccc acaagcgcat gaggactgtc accaactact
     2101 tccttgtgaa cctggctttc tccgacgcct ccatggccgc cttcaacacg ttggtcaatt
     2161 tcatctacgc gcttcatagc gagtggtact ttggcgccaa ctactgccgc ttccagaact
     2221 tctttcctat cacagctgtg ttcgccagca tctactccat gacggccatt gcggtggaca
     2281 ggtatatggc tattattgat cccttgaaac ccagactgtc tgctacagca accaagattg
     2341 tcattggaag tatttggatt ctagcatttc tacttgcctt ccctcagtgt ctttattcca
     2401 aaaccaaagt catgccaggc cgtactctct gctttgtgca atggccagaa ggtcccaaac
     2461 aacatttcac ttaccatatt atcgtcatta tactggtgta ctgtttccca ttgctcatca
     2521 tgggtattac atacaccatt gttggaatta ctctctgggg aggagaaatc ccaggagata
     2581 cctgtgacaa gtatcatgag cagctaaagg ccaaaagaaa ggttgtcaaa atgatgatta
     2641 ttgttgtcat gacatttgct atctgctggc tgccctatca tatttacttc attctcactg
     2701 caatctatca acaactaaat agatggaaat acatccagca ggtctacctg gctagctttt
     2761 ggctggcaat gagctcaacc atgtacaatc ccatcatcta ctgctgtctg aataaaagat
     2821 ttcgagctgg cttcaagaga gcatttcgct ggtgtccttt catcaaagtt tccagctatg
     2881 atgagctaga gctcaagacc accaggtttc atccaaaccg gcaaagcagt atgtacaccg
     2941 tgaccagaat ggagtccatg acagtcgtgt ttgaccccaa cgatgcagac accaccaggt
     3001 ccagtcggaa gaaaagagca acgccaagag acccaagttt caatggctgc tctcgcagga
     3061 attccaaatc tgcctccgcc acttcaagtt tcataagctc accctatacc tctgtggatg
     3121 aatattctta agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     3181 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     3241 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     3301 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     3361 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     3421 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     3481 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     3541 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     3601 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     3661 agatccaagc tgtgaccggc gcctacgcta gacgccacca tgaccgagta caagcccacg
     3721 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3781 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3841 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3901 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3961 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4021 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4081 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4141 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4201 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4261 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa
     4321 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4381 gctgctttaa tgcctttgta tcagttaact tgtttattgc agcttataat ggttacaaat
     4441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4501 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat
     4561 tagtctatca gaagctatct ggtctccctt ccgggggaca agacatccct gtttaatatt
     4621 taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa ccaggttgca
     4681 gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc
     4741 ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc ccgtatcccc
     4801 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     4861 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     4921 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     4981 tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag aaaatatata
     5041 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     5101 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     5161 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     5221 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     5281 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     5341 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     5401 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     5461 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     5521 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     5581 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     5641 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     5701 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     5761 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     5821 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5881 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5941 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6001 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6061 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6121 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6181 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6241 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6301 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6361 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6421 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6481 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6541 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6601 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6661 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6721 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6781 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6841 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6901 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6961 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7021 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7081 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7141 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7201 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7261 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7321 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7381 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7441 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7501 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7561 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7621 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7681 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7741 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7801 gggaataagg gcgacacgga aatgttgaat actcat
//
