LOCUS       V025607                 8157 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025607
VERSION     V025607
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..2862
                     /label="MINDY2(NM_001040450)"
                     /note="MINDY2(NM_001040450)"
                     /gene="MINDY2"
     polyA_signal    2916..3538
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3580..3877
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3931..4725
                     /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    5377..5511
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(5517..5535)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5549..5565)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5573..5589
                     /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(5597..5627)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5642..5663
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5951..6539)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6710..7570)
                     /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(7571..7675)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7702..8130
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tggagagcag ccccgagagc
     1021 ctgcagccgc tagaacacgg ggtggcggcc gggccagcgt cagggacagg ttcttcgcag
     1081 gaagggctac aggagaccag gctcgccgct ggtgatggtc ctggggtatg ggcggcggag
     1141 accagcggcg ggaatgggct gggggcggcg gccgccagga ggagcctccc ggactcggct
     1201 tctcccgcgg gctctcctga ggttcccgga ccctgcagct cctccgcggg tttggacttg
     1261 aaggacagtg gtttggagag tcctgctgcc gccgaggcgc ctctgagagg gcagtacaag
     1321 gtgaccgcct ccccggagac agccgtggcc ggagtgggtc atgagttggg taccgccgga
     1381 gacgcgggag cccgcccgga tctcgccggc acctgccaag cagaactgac cgccgccggc
     1441 tccgaagagc ccagcagcgc cggcggcctc agcagcagtt gcagcgaccc gagccctcct
     1501 ggggaatctc cgagcctgga ctctctggag tcgttctcta acctgcattc ttttcccagt
     1561 agctgcgagt tcaatagtga ggagggagcg gagaacaggg tccctgagga ggaggagggc
     1621 gcggcggtgt tgcccggggc tgttcctctg tgcaaggagg aggaggggga ggagaccgct
     1681 caggtgctgg cggcctccaa ggaacgcttc ccgggacaat ctgtgtatca catcaagtgg
     1741 atccagtgga aggaagagaa cacacccatc atcacccaga atgagaacgg accctgcccc
     1801 ttgctggcca tcctcaatgt tttgctcctg gcctggaagg tgaaacttcc accgatgatg
     1861 gaaatcataa ctgctgagca gctgatggaa tatttaggag attacatgct tgatgcaaag
     1921 ccaaaagaaa tttcagaaat tcaacgttta aattatgaac agaatatgag tgatgccatg
     1981 gcaattttgc acaaactaca gacaggcctg gatgtaaatg taagattcac tggtgttcga
     2041 gtgtttgaat atacaccaga atgcatagta tttgatcttc ttgatattcc tttgtaccat
     2101 gggtggttag tagaccctca gattgatgac attgtaaaag ctgttggtaa ctgcagctac
     2161 aaccaactag tggagaagat catctcttgt aaacagtcag acaatagtga gctggttagt
     2221 gaaggctttg tagctgagca gtttctaaat aacacagcca ctcaactgac ataccatgga
     2281 ttatgtgaac taacttcaac ggttcaggaa ggagaacttt gtgtgttctt tcggaataat
     2341 cattttagca ccatgaccaa atacaagggt caactgtatt tgttggtaac ggaccagggg
     2401 tttcttactg aagagaaagt tgtttgggaa agcctacaca acgtagatgg tgatggaaat
     2461 ttctgtgact cagaatttca tcttcgacct ccttcagatc ctgaaactgt atacaaagga
     2521 caacaagatc agatagatca ggattatctt atggcattat ctctacaaca agaacagcag
     2581 agccaagaga tcaattggga acaaatcccg gaaggaatca gtgatttgga actagcaaag
     2641 aaactccaag aggaagagga cagacgggct tctcaatact atcaggaaca ggaacaagca
     2701 gcagctgctg ctgctgctgc ttctacacag gctcagcagg gccagccagc acaagcctct
     2761 ccatcaagtg gaagacaatc tgggaatagt gaacgtaaac ggaaggaacc acgagaaaaa
     2821 gataaagaaa aagaaaagga aaaaaatagc tgtgttattt tgtaagaatt catcgataga
     2881 tctgatatcg gtaccagtcg actctagagg atcccgggtg gcatccctgt gacccctccc
     2941 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     3001 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     3061 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     3121 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     3181 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     3241 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     3301 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     3361 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     3421 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     3481 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     3541 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg aattgatcag cttgggatct
     3901 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3961 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4021 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4081 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4141 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4201 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4261 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4321 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4381 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4441 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4501 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4561 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4621 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4681 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4741 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4801 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4861 agcgcgggga tctcatgctg gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt
     4921 ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg
     4981 tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt
     5041 ggggaggcac gatggccgct ttggtcgacc cggacgggac gctcctgcgc ctgatacaga
     5101 acgaattgct tgcaggcatc tcatgagtgt gtcttcccgt tttccgcctg aggtcactgc
     5161 gtggatggag cgctggcgcc tgctgcgcga cggcgagctg ctcaccaccc actcgccaag
     5221 ctggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc gatcataatc
     5281 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     5341 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     5401 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5461 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caattcccta
     5521 tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     5581 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5641 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5701 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5761 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5821 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5881 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5941 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6001 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6061 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6121 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6181 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6241 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6301 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6361 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6421 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6481 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6541 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6601 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6661 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6721 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6781 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6841 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6901 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6961 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7021 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7081 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7141 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7201 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7261 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7321 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7381 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7441 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7501 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7561 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7621 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7681 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7741 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     7801 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7861 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     7921 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7981 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8041 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8101 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaattt
//
