LOCUS       V021730                 8744 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021730
VERSION     V021730
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..3448
                     /label="Atg9b(NM_001002897)"
                     /note="Atg9b(NM_001002897)"
                     /gene="Atg9b"
     CDS             3455..3496
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    3520..4108
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4174..4222
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4424..4852
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4866..5195
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5262..6056
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6232..6353
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6402..6418)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6426..6442
                     /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(6450..6480)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6495..6516
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6804..7392)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7563..8423)
                     /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"
     promoter        complement(8424..8528)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggtgag acgaacgggg tgggggggga gcaggaggca
      721 acgggggcgg tggggagact tggggcccag ctcagtgcct ctgctcccta tggcactgcc
      781 actgcctgct tctccatgcc ggggaactgg gggaaggagg atctctgtct tctctctgtc
      841 ccctgcccct cgcacaagaa gctgctcctc ttcagtgttc cctccggcct cagggtctcc
      901 ctgcctggtg attcaggagg caggggcttc tcagactcca cacaatgtct tacccacccc
      961 gaccacaccc tcaacgcagg cccatcccac aatgatacac acctctgctt ccccctcgtg
     1021 ggggtcccac tccaccccac ccctggcctc ggccactccc cctccctcct gcccgcgccc
     1081 ccaggaccat cctgggctgc ggatgggccc tctgatccct gagcaggact atgaacggct
     1141 agaggactgt gaccctgagg ggtctcaaga ctcccctatc catggggagg accaccaacc
     1201 cttgcttcat gtacccgaag gactccgcgg ttcctggcat cacatccaga acctggacag
     1261 ttttttcacc aagatctaca gctatcatca gcggaatggc tttgcctgta tcctgctgga
     1321 ggatgtcttc cagctggggc agttcatttt cattgtcacc tttacaacct tcctccttcg
     1381 ctgcgtggat tacaatgttc tcttcaacaa ccaaccaaag aaccatacaa gacgtggacc
     1441 attacacagc aaagtgacct tgtcagatgc cattctaccc tcggcccagt gtgcagagaa
     1501 gatccacgac agtcccctgt tggtcttcct cctagtcctg gctgctggct tctggctgtt
     1561 ccagctgctt cgctcggtct gcaacctctt cagctactgg gacatccagg tgttttacag
     1621 ggaggccctg cacattcccc cagaggagct cagctcggta ccctgggccg aggtgcagtc
     1681 tcgcctcctg gaactgcaga ggagcggtgg gctgtgtgtg cagccgcggc cgctgacgga
     1741 actggacgtc caccaccgca tcctgcgcta caccaactac caggtagcgt tagccaacaa
     1801 gggtctgctg cctgcccgct gcccgctgcc ctgggggagc agcgccgcct tcctcagccg
     1861 cggcctggcg ctcaacgtgg acttgctgct cttccgcggt cccttctcac tcttccgcgg
     1921 gggctgggag cttcccgaag cctacaagcg cagcgacctc cggggcgtcc tggctaaccg
     1981 ctggcggcgc acggtgctgc tgctggccgc cgtgaacttg gccctgagcc cgctggtgct
     2041 ggcctggcag gtattgcacg ccttctacag ccacgtggag ctgctgcggc gcgagcccgg
     2101 agccttcggg gcgcgccgct ggtcccgcct ggcccgcctg cagcttcgcc actttaacga
     2161 gctgccgcac gagctgcgtg cgcgcctggg ccgcgcctac cggcccgccg ccgccttcct
     2221 gcgtgcagcg gagcccccag ctcccctgcg cgcgctgctg gcccgccagc tggtcttctt
     2281 ctcgggagct ctctttgccg cgctgctagt gctcaccatc tacgatgagg acgtgttggc
     2341 cgtggagcat gtgctcacta ccatgacggc gctcggggtc acagccactg tggccaggtc
     2401 cttcattcca gaagagcaat gccaggggcg ttcctcccag cttctgctgc aggcagccct
     2461 ggcgcacatg cattacctcc cagaggagcc tggtgcaacg ggtgctcgag ccagctctta
     2521 ctggcagatg gcgcagctgc ttcagtaccg agcagtctcc ctcctggaag agctcctgtc
     2581 tccactcctg accccattgt ttctgctctt ctggttccgc ccccgtgctc tggagattat
     2641 tgactttttt catcacttca ctgtggatgt ggctggtgta ggggacatct gttcttttgc
     2701 attaatggat gtgaagcgcc acgggcatcc tcagtggctt tcagagggac agacagaggc
     2761 ctcactctct cagcgtgcag aggatggaaa gaccgaactc tccttaatgc ggttctccct
     2821 ggcacaccca caatggcagc caccagggca cagctctaag ttccttggac accttcgggg
     2881 ccgggtacaa caagatgcag ctgcctgggg tgctccctcg actcggagtc cccccacccc
     2941 aggagtgctc agtgactgta catcccctct gccagaagcc ttccttgcca acctcttggt
     3001 aaatccccgc ccaccccaga gggacctgag tcccacagcc ccctgccctg ctgcagccac
     3061 agccagcctc cttgcctcta tttcacgaat ggtccaggac cccagctgtg tgtccccagg
     3121 aggcacaggg ggccagaagc tgacccagct cccagagctt gtctctgctg agatgagcct
     3181 ccacgctatt tacctacatc agcttcatca acagcagcag caagaactat ggggagaggc
     3241 ttcagcttcc tccccatcca ggccctggtc cagcccttca cagccaggct cacctgatga
     3301 ggagaagcca tcttggtcaa gtgatggctc cagtcctgct tctagcccaa gacaacagtg
     3361 gggcacccag agggcccaga atctgttccc aaaggggttt caggagaaca cagacaccca
     3421 gaaggagccc ctaacaggcc ccttgcacct cgagggtaag cctatcccta accctctcct
     3481 cggtctcgat tctacgtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     3541 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3601 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3661 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3721 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3781 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3841 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3901 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3961 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4021 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4081 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     4141 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4201 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4261 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4321 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4381 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4441 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4501 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4561 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     4621 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4681 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4741 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     4801 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     4861 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4921 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4981 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5041 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5101 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     5161 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     5221 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     5281 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     5341 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     5401 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     5461 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     5521 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     5581 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     5641 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     5701 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     5761 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     5821 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5881 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5941 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6001 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6061 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6121 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6181 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6241 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     6301 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6361 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6421 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6481 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6541 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6601 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6661 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6721 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6781 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6841 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6901 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6961 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     7021 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7081 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7141 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7201 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     7261 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7321 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7381 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7441 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7501 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7561 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7621 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7681 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7741 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7801 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7861 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7921 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7981 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8041 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8101 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8161 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8221 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8281 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8341 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8401 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8461 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8521 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     8581 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     8641 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     8701 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
