LOCUS       V021243                 7154 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021243
VERSION     V021243
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..1801
                     /label="AIFM2(NM_001198696)"
                     /note="AIFM2(NM_001198696)"
                     /gene="AIFM2"
     CDS             1841..1906
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1930..2518
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2584..2632
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2834..3262
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3276..3605
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3672..4466
                     /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    4642..4763
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4812..4828)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4836..4852
                     /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(4860..4890)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4905..4926
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5214..5802)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5973..6833)
                     /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(6834..6938)
                     /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 ccatggggtc ccaggtctcg gtggaatcgg gagctctgca
      721 cgtggtgatt gtgggtgggg gctttggcgg gatcgcagca gccagccagc tgcaggccct
      781 gaacgtcccc ttcatgctgg tggacatgaa ggactccttc caccacaatg tggctgctct
      841 ccgagcctcc gtggagacag ggttcgccaa aaagacattc atttcttact cggtgacttt
      901 caaggacaac ttccggcagg ggctagtagt ggggatagac ctgaagaacc agatggtgct
      961 gctgcagggt ggcgaggccc tgcccttctc tcatcttatc ctggccacgg gcagcactgg
     1021 gcccttcccg ggcaagttta atgaggtttc cagccagcag gccgctatcc aggcctatga
     1081 ggacatggtg aggcaggtcc agcgctcacg gttcatcgtg gtggtgggag gaggctcggc
     1141 tggagtggag atggcagcag agattaaaac agaatatcct gagaaagagg tcactctcat
     1201 tcactcccaa gtggccctgg ctgacaagga gctcctgccc tccgtccggc aggaagtgaa
     1261 ggagatcctc ctccggaagg gcgtgcagct gctgctgagt gagcgggtga gcaatctgga
     1321 ggagctgcct ctcaatgagt atcgagagta catcaaagtg cagacggaca aaggcacaga
     1381 ggtggccacc aacctggtga ttctctgcac cggcatcaag atcaacagct ccgcctaccg
     1441 caaagcgttt gagagcagac tagccagcag tggtgctctg agagtgaacg agcacctcca
     1501 ggtggagggc cacagcaacg tctacgccat tggtgactgt gccgacgtga ggacgcccaa
     1561 gatggcctat cttgccggcc tccacgccaa catcgccgtg gccaacatcg tcaactctgt
     1621 gaagcagcgg cctctccagg cctacaagcc gggtgcactg acgttcctcc tgtccatggg
     1681 gagaaatgac ggtgtgggcc aaatcagtgg cttctatgtg ggccggctca tggttcggct
     1741 gaccaagagc cgggacctgt tcgtctctac gagctggaaa accatgaggc agtctccacc
     1801 tgaattctgc agatatccag cacagtggcg gccgctcgag gactacaaag accatgacgg
     1861 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     1921 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1981 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2041 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2101 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2161 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2221 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2281 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2341 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2401 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2461 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2521 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     2581 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     2641 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2701 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     2761 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     2821 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3181 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     3241 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3361 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3421 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3481 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3541 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3601 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3661 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3721 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3781 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3841 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3901 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3961 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4021 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4081 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4141 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4201 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4261 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4321 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4381 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4441 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4501 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4561 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4621 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4681 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4741 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4801 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4861 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4921 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4981 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5041 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5401 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5641 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5761 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5881 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5941 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6001 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6061 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6121 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6181 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6241 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6301 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6361 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6421 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6481 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6541 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6601 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6661 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6721 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6781 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6841 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6901 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6961 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7021 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     7081 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     7141 agaatctgct tagg
//
