LOCUS       V043898                 8443 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043898
VERSION     V043898
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        1..1179
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     promoter        1196..1214
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     protein_bind    1293..1317
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             1347..4250
                     /label="Aars1(NM_146217)"
                     /note="Aars1(NM_146217)"
                     /gene="Aars1"
     CDS             4257..4283
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4287..4313
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             4317..4343
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4371..4595
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4665..4698
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
     primer_bind     complement(5156..5172)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        5646..5750
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5751..6611
                     /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"
     rep_origin      6782..7370
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7658..7679
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7694..7724
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7732..7748
                     /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"
     primer_bind     7756..7772
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8270..8303
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
                     /label="loxP"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
     1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
     1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
     1321 cggccgcccc cttcaccatg gccaccatgg atgccacttt aacagcacga gagatccgag
     1381 agcggtttat aaatttcttc agacgaaatg agcacaccta tgtccactct tccgccacca
     1441 tccctctgga tgaccccact ctgctcttcg ccaacgctgg catgaaccag ttcaaaccca
     1501 tcttcctaaa cacaatcgac ccatctcacc ctatggcgaa gctgagcaga gctgccaata
     1561 cccagaaatg catccgagct gggggcaagc acaacgacct ggatgacgtg ggcaaggatg
     1621 tctaccacca cactttcttt gagatgctgg gctcctggtc ttttggggac tatttcaagg
     1681 aattggcatg taagatggcc ctggaacttc ttacccagga gtttggtatt ccagttgaaa
     1741 gactctatgt cacttacttt ggtggtgatg aggcggccgg cctagagcca gatctggagt
     1801 gcagacagat ttggcaaaat ttaggactgg atgaagccag aatcctccct ggcaacatga
     1861 aggacaactt ctgggagatg ggtgacacag gcccctgtgg tccctgcagc gagatccact
     1921 atgaccgcat tggtggcagg gacgccgcac accttgtcaa ccaggatgac cccaatgtgc
     1981 tggagatctg gaacctcgtg ttcatccagt ataacaggga gtcggatggt gttctgaaac
     2041 ctcttcccaa gaaaagcatc gacacaggga tgggcctaga gaggctggtg tctgtgctgc
     2101 agaacaagat gtccaactac gacaccgacc ttttcatgcc ttacttcgaa gccattcaga
     2161 agggcaccgg cgcccggccg tatactggga aggttggtgc tgaggatgca gacggaatcg
     2221 acatggccta ccgggttctg gctgaccacg cccggaccat cactgtggcg ctggctgatg
     2281 gcggccggcc tgacaacaca ggacgggggt acgtgctgag acggatcctt cgccgagctg
     2341 ttcggtattc ccatgagaaa ctgaatgcca gcaggggttt cttcgctaca ttggttgatg
     2401 ttgtcgttca atccctggga gacgcatttc ctgagctgaa gaaggaccca gagatggtga
     2461 aggacatcat taatgaagaa gaggtgcagt ttctcaagac tctcagccga gggcggcgca
     2521 tcctggaccg aaaaattcag agcttaggag actgcaaaac catcccaggt gacaccgctt
     2581 ggctcctcta tgatacttat ggattcccag tggacctcac tggattgatt gctgaagaga
     2641 agggcctggt ggtagatatg aatggctttg aggaagagag gagactggcc cagctgaagt
     2701 cacagggcaa gggagcaggg gacgaagacc tcatcatgct ggacatttac gctattgaag
     2761 agctccgggc aaagggtttg gaggccacag atgattctcc aaagtataac taccaatcag
     2821 actccagtgg tagctacgtg tttgagtgta cagtggctac cgtgcttgct ctgcgtcggg
     2881 aaaagatgtt tgtggacgag gtggtcactg gccaggagtg tggagtggtg ctggacaaga
     2941 cctgtttcta tgcggagcaa ggagggcaga tctacgatga aggctacttg gtgaaggtcg
     3001 acgacagcag tgaggacaaa accgagttta cagtgaagaa cgctcaggtc cgaggcgggt
     3061 acgtgctgca cataggaacg atatatggta acctgaaagt gggggatcaa gtccggctgt
     3121 ttattgatga gccccggcgg agacccgtca tgagcaacca cacagctaca cacatcctca
     3181 acttcgcctt gcgctcggtg cttggggaag cggaccagaa aggttcactg gttgctcctg
     3241 accgccttcg gtttgacttc actgccaagg gagccatgtc cacccagcaa atcaagaagg
     3301 ctgaagagat tgtcaatggc atgattgagg cagctaagcc tgtctacacc caggattgtc
     3361 ccctggcagc agcaaaagcc atccagggcc ttcgagctgt gtttgatgag acctatcctg
     3421 accccgttcg agtcgtctcc attggggtcc ccgtgtcaga gctgctggat gacccgtgtg
     3481 ggcctgctgg ctcccttact tctgttgagt tctgtggagg aacacatcta cggaattcta
     3541 gccacgcagg cgcttttgtg atagtgacag aagaagctat tgccaagggt atccggagga
     3601 ttgttgctgt cacaggtgct gaagcccaga aggctctcag gaagtcagaa accttgaaga
     3661 aatctctctc tgccatggag gccaaagtga aggcccagac tgcacccaac aaggacgtgc
     3721 agagggaaat tgctgacctc ggtgaggccc tggccactgc agtcatcccc cagtggcaga
     3781 aagatgaaca acgagagact ctgaagtccc tgaagaaagt catggatgac ctagaccggg
     3841 ccagcaaagc cgatgtgcag aagcgagtgt tagagaagac aaagcagctg attgacagca
     3901 accccaacca gccgctggtc atcctggaga tggagagtgg agcatcggcc aaggccctga
     3961 atgaagcgtt gaagcttttc aagacgcatt ctcctcagac atctgccatg ctgttcacgg
     4021 tggacaatga ggctggcaag atcacgtgct tgtgccaggt tccccagaat gcagccaacc
     4081 ggggactgaa agccagcgag tgggtccagc aggtttcagg cctgatggat ggtaaaggcg
     4141 gtggcaagga catgtccgcc caggccacag gcaagaacgt gggctgcctt caggaagcac
     4201 tgcagctggc tacctccttt gcccagctcc gtctgggaga tgtgaagaac ctcgagtacc
     4261 catacgacgt acctgattat gcaacttatc cttatgatgt acctgattat gctagctatc
     4321 cttatgatgt cccagactac gcatagttta aacccgctga tcagcctcga ctgtgccttc
     4381 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4441 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4501 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4561 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4621 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaaataact tcgtataatg
     4681 tatgctatac gaagttatat cgtatgcatg ctttgcatac ttctgcctgc tggggagcct
     4741 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttccttgaaa tacatccaca
     4801 ggtacacctc caattgactc aaatgatgtc aattagtcta tcagaagctt ctaaagccat
     4861 gacatcattt tctggaattt tccaagctgt ttaaaggcac agtcaactta gtgtatgtaa
     4921 acttctgacc cactggaatt gtgatacagt gaattataag tgaaataatc tgtctgtaaa
     4981 caattgttgg aaaaatgact tgtgtcatgc acaaagtaga tgtcctaact gacttgccaa
     5041 aactattgtt tgttaacaag aaatttgtgg agtagttgaa aaacgagttt taatgactcc
     5101 aacttaagtg tatgtaaact tccgacttca actgtagggt accgagctcg aattcactgg
     5161 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     5221 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     5281 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     5341 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     5401 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5461 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5521 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5581 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5761 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5821 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5881 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5941 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6001 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6061 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6121 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6181 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6241 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6301 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6361 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6421 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6481 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6541 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6601 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6661 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6721 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6781 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6841 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6901 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6961 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7021 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7081 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7141 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7201 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7261 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7321 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7381 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7441 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7501 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7561 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7621 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7681 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7741 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc
     7801 aggtcgactc tagacctata cagttgaagt cggaagttta catacactta agttggagtc
     7861 attaaaactc gtttttcaac tactccacaa atttcttgtt aacaaacaat agttttggca
     7921 agtcagttag gacatctact ttgtgcatga cacaagtcat ttttccaaca attgtttaca
     7981 gacagattat ttcacttata attcactgta tcacaattcc agtgggtcag aagtttacat
     8041 acactaagtt gactgtgcct ttaaacagct tggaaaattc cagaaaatga tgtcatggct
     8101 ttagaagctt ctgatagact aattgacatc atttgagtca attggaggtg tacctgtgga
     8161 tgtatttcaa ggggatccag acatgataag atacattgat gagtttggac aaaccacaac
     8221 tagaatgcag cggccgctag ctcgagatct taattaaact agtatcgata taacttcgta
     8281 taatgtatgc tatacgaagt tatatgtacg ggccagatat acgcgttgac attgattatt
     8341 gactaggctt ttgcaaaaag ctttgcaaag atggataaag ttttaaacag agaggaatct
     8401 ttgcagctaa tggaccttct aggtcttgaa aggagtggga att
//
