LOCUS       V021975                 8723 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021975
VERSION     V021975
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             1722..3542
                     /label="ATG16L1(NM_030803)"
                     /note="ATG16L1(NM_030803)"
                     /gene="ATG16L1"
     CDS             3582..4301
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     promoter        4366..4577
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     CDS             4587..5186
                     /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    5294..5415
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6702..6718)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6726..6742
                     /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(6750..6780)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6795..6816
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7104..7692)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7863..8723)
                     /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 ctagagccac catgtcgtcg ggcctccgcg
     1741 ccgctgactt cccccgctgg aagcgccaca tctcggagca actgaggcgc cgggaccggc
     1801 tgcagagaca ggcgttcgag gagatcatcc tgcagtataa caaattgctg gaaaagtcag
     1861 atcttcattc agtgttggcc cagaaactac aggctgaaaa gcatgacgta ccaaacaggc
     1921 acgagataag tcccggacat gatggcacat ggaatgacaa tcagctacaa gaaatggccc
     1981 aactgaggat taagcaccaa gaggaactga ctgaattaca caagaaacgt ggggagttag
     2041 ctcaactggt gattgacctg aataaccaaa tgcagcggaa ggacagggag atgcagatga
     2101 atgaagcaaa aattgcagaa tgtttgcaga ctatctctga cctggagacg gagtgcctag
     2161 acctgcgcac taagctttgt gaccttgaaa gagccaacca gaccctgaag gatgaatatg
     2221 atgccctgca gatcactttt actgccttgg agggaaaact gaggaaaact acggaagaga
     2281 accaggagct ggtcaccaga tggatggctg agaaagccca ggaagccaat cggcttaatg
     2341 cagagaatga aaaagactcc aggaggcggc aagcccggct gcagaaagag cttgcagaag
     2401 cagcaaagga acctctacca gtcgaacagg atgatgacat tgaggtcatt gtggatgaaa
     2461 cttctgatca cacagaagag acctctcctg tgcgagccat cagcagagca gccactaagc
     2521 gactctcgca gcctgctgga ggccttctgg attctatcac taatatcttt gggagacgct
     2581 ctgtctcttc cttcccagtc ccccaggaca atgtggatac tcatcctggt tctggtaaag
     2641 aagtgagggt accagctact gccttgtgtg tcttcgatgc acatgatggg gaagtcaacg
     2701 ctgtgcagtt cagtccaggt tcccggttac tggccactgg aggcatggac cgcagggtta
     2761 agctttggga agtatttgga gaaaaatgtg agttcaaggg ttccctatct ggcagtaatg
     2821 caggaattac aagcattgaa tttgatagtg ctggatctta cctcttagca gcttcaaatg
     2881 attttgcaag ccgaatctgg actgtggatg attatcgatt acggcacaca ctcacgggac
     2941 acagtgggaa agtgctgtct gctaagttcc tgctggacaa tgcgcggatt gtctcaggaa
     3001 gtcacgaccg gactctcaaa ctctgggatc tacgcagcaa agtctgcata aagacagtgt
     3061 ttgcaggatc cagttgcaat gatattgtct gcacagagca atgtgtaatg agtggacatt
     3121 ttgacaagaa aattcgtttc tgggacattc gatcagagag catagttcga gagatggagc
     3181 tgttgggaaa gattactgcc ctggacttaa acccagaaag gactgagctc ctgagctgct
     3241 cccgtgatga cttgctaaaa gttattgatc tccgaacaaa tgctatcaag cagacattca
     3301 gtgcacctgg gttcaagtgc ggctctgact ggaccagagt tgtcttcagc cctgatggca
     3361 gttacgtggc ggcaggctct gctgagggct ctctgtatat ctggagtgtg ctcacaggga
     3421 aagtggaaaa ggttctttca aagcagcaca gctcatccat caatgcggtg gcgtggtcgc
     3481 cctctggctc gcacgttgtc agtgtggaca aaggatgcaa agctgtgctg tgggcacagt
     3541 acggtggttc tggtggtggt tctggtggac cggtcgccac catggtgagc aagggcgagg
     3601 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     3661 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     3721 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     3781 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     3841 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     3901 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     3961 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4021 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4081 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4141 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4201 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4261 ccgccgggat cactctcggc atggacgagc tgtacaagta agaattcgaa tttaaatcgg
     4321 atccgcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     4381 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     4441 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     4501 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4561 ttgccgccag aacacagctg aagcttatga ccgagtacaa gcccacggtg cgcctcgcca
     4621 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     4681 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     4741 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     4801 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     4861 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     4921 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     4981 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     5041 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     5101 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     5161 gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat
     5221 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     5281 ctttgtatca gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5341 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5401 catcaatgta tcttatcatg tctggaattg actcaaatga tgtcaattag tctatcagaa
     5461 gctatctggt ctcccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt
     5521 tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc
     5581 tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg gaattgactg
     5641 gattcctttt ttagggccca ttggtatggc tttttccccg tatcccccca ggtgtctgca
     5701 ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc
     5761 ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg
     5821 agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg
     5881 gctttggggg ggggctgtcc ctgatatcta taacaagaaa atatatatat aataagttat
     5941 cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca
     6001 cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca aaatcagtga
     6061 cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta
     6121 aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg
     6181 cgtcaatttt acgcagacta tctttctagg gttaatctag ctgcatcagg atcatatcgt
     6241 cgggtctttt ttccggctca gtcatcgccc aagctggcgc tatctgggca tcggggagga
     6301 agaagcccgt gccttttccc gcgaggttga agcggcatgg aaagagtttg ccgaggatga
     6361 ctgctgctgc attgacgttg agcgaaaacg cacgtttacc atgatgattc gggaaggtgt
     6421 ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc acctgggata ccagttcgtc
     6481 gcggcttttc cggacacagt tccggatggt cagcccgaag cgcatcagca acccgaacaa
     6541 taccggcgac agccggaact gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct
     6601 gggatattac gtcagcgagg acgggtatcc tggctggatg ccgcagaaat ggacatggat
     6661 accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     6721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7501 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8701 acacggaaat gttgaatact cat
//
