LOCUS       V044003                 8428 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044003
VERSION     V044003
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3918
                     /label="AARS2(NM_020745)"
                     /note="AARS2(NM_020745)"
                     /gene="AARS2"
     CDS             3925..3990
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4028..4252
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4298..4726
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4740..5069
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5136..5930
                     /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    6104..6225
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6274..6290)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6298..6314
                     /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(6322..6352)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6367..6388
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6676..7261)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7432..8292)
                     /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(8293..8397)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcag cgtcagtggc agctgcagcc cggaggctgc ggcgggccat tcgaaggtcg
     1021 cccgcatggc ggggcctcag ccatcggccg ctctcatcgg agccccctgc agccaaggcc
     1081 tcggccgtga gggccgcctt tctgaacttc tttcgggacc gccatggcca ccggctggtg
     1141 ccctccgctt ccgtgcggcc ccgcggcgac cccagtttgc tttttgtcaa tgcgggcatg
     1201 aaccagttca agccaatctt tctgggcacc gtggatccac gaagcgagat ggcaggcttc
     1261 cgacgtgtgg ccaacagcca gaaatgtgtg agagctggag gacaccataa cgacctggaa
     1321 gatgtgggtc gagacctttc ccatcatacc ttctttgaaa tgcttggcaa ttgggccttt
     1381 gggggtgaat attttaagga ggaggcttgt aacatggcct gggaactgct gactcaggtc
     1441 tatgggatcc ctgaggaaag gctctggatc tcctactttg atggtgaccc caaggcaggg
     1501 ctggacccag acctggagac cagggacatc tggctgagct taggggtgcc tgctagccgt
     1561 gtgctttcct ttggaccaca agagaacttc tgggagatgg gggatactgg cccttgtggg
     1621 ccctgtactg agatccacta cgaccttgct ggtggggtgg gagcccccca gctggtagag
     1681 ctttggaacc tggtcttcat gcaacacaac agagaggcag atggaagcct gcagcccctg
     1741 ccccagcggc atgtggacac aggaatgggc ctggaaaggc tggtggctgt gctgcaaggc
     1801 aaacactcca cctatgacac tgacctcttt tccccgctgc tcaacgccat acagcagggc
     1861 tgcagggcac ccccttactt gggccgagta ggggtggcag acgaggggcg cacagacaca
     1921 gcgtaccgcg tggtggctga ccacatccgc acactcagtg tctgcatctc tgatggcatc
     1981 ttccctggga tgtcaggtcc cccgctggtt cttcgtcgga tcctgcgtcg agctgtgcgt
     2041 ttctccatgg agatcttaaa ggcaccacct ggcttcctag gcagcctggt acctgtagtg
     2101 gtggagacac tgggagatgc ttatccagaa ctgcaaagga actcagccca gatcgccaac
     2161 ctggtgtcag aggacgaggc agccttcctg gcctccctgg agcggggtag gcggatcatt
     2221 gatcggactc tgaggaccct ggggccttca gatatgttcc ctgctgaagt ggcctggtcc
     2281 ttgtcactgt gtggagacct gggactcccc ttggacatgg tagagctgat gctggaggag
     2341 aaaggggtcc agctagactc cgctggactg gagcggttgg cccaagagga ggcccagcac
     2401 cgggcacggc aggctgagcc agttcagaag cagggattgt ggcttgatgt ccatgcgctt
     2461 ggggagctgc agcgccaagg agtgccccca actgacgaca gccccaagta caactactcc
     2521 ctgcgaccca gcggaagtta tgagttcggc acctgtgagg cccaggtgtt gcaactgtat
     2581 acagaggacg ggacagcagt ggcctccgtg gggaaaggcc agcgctgtgg cctcctcttg
     2641 gacaggacca acttctacgc agaacagggg ggccaggctt cagaccgtgg ctacctggtg
     2701 cgggcagggc aagaggacgt gctgttccca gtagcccggg cccaggtctg tggaggtttc
     2761 atcctgcatg aggcagtagc ccctgagtgc ctgcggttag gggaccaggt gcagctgcat
     2821 gtggatgagg cctggcgtct aggctgcatg gcgaagcata cggccaccca cctgctgaac
     2881 tgggcactga ggcagaccct gggccctggc acagagcagc agggctccca tctcaatcct
     2941 gagcagctgc gcttggatgt gaccacccag accccattga ccccagagca gctccgggca
     3001 gtggagaaca ctgtgcagga ggccgtgggg caggatgagg ctgtgtacat ggaggaggtg
     3061 cccctggcgc tcactgccca ggtccctggc ctgcgctctc tggatgaggt ttacccagac
     3121 cctgtgcggg tggtatcagt gggggtgccc gtggcccatg cattggaccc agcctcccaa
     3181 gccgcactgc agacctctgt ggagctatgc tgtgggacgc acctgttacg tactggggct
     3241 gtaggggacc tggttatcat cggggaccgc cagctttcca agggcactac ccgcctgctg
     3301 gccgtcactg gggagcaggc ccagcaggcc cgagagctag gccagagcct ggcccaggaa
     3361 gtgaaagcgg ccactgagcg gctgagtctg gggagccggg atgtggcgga ggcactgagg
     3421 ctgtccaagg acataggacg actcattgaa gctgtggaaa ctgctgtgat gccccagtgg
     3481 cagcggcggg agctgctggc cacagtgaag atgctgcagc ggcgtgccaa cactgccatc
     3541 cgtaagctgc aaatgggaca ggctgcaaag aaaactcagg agctgctgga gcggcactcg
     3601 aaggggcctc tgattgtgga cacagtctct gctgagtctc tctcagtgct ggtgaaggtg
     3661 gtacggcagc tgtgtgagca ggcccccagc acgtctgtgc tcctactcag cccccagccc
     3721 atggggaagg tgctgtgtgc ctgtcaggtg gcccagggtg ccatgcccac cttcacagca
     3781 gaggcctggg cactggcagt gtgcagccac atggggggca aggcgtgggg ctcacgagtg
     3841 gtggcccaag gcaccggaag cactactgac ctggaagctg ccctcagtat agcccaaacc
     3901 tatgccctca gccagctcct cgaggactac aaagaccatg acggtgatta taaagatcat
     3961 gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac ccgctgatca
     4021 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4201 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4261 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4321 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4381 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4441 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4501 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4561 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4621 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4681 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4741 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4801 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4861 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4921 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4981 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5041 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5101 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5161 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5221 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5281 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5341 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5401 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5461 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5521 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5581 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5641 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5701 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5761 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5821 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5881 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5941 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6001 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6061 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6121 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6181 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6241 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7201 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8401 atttccccga aaagtgccac ctgacgtc
//
