LOCUS       V019033                 8528 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019033
VERSION     V019033
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             1324..4398
                     /label="NAT10(NM_024662)"
                     /note="NAT10(NM_024662)"
                     /gene="NAT10"
     CDS             4399..4422
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    4456..4680
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     protein_bind    4750..4783
                     /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(5241..5257)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        5731..5835
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             5836..6696
                     /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      6867..7455
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    7743..7764
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        7779..7809
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7817..7833
                     /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     7841..7857
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    8355..8388
                     /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 gcaggctgcc
     1321 accatgcatc ggaaaaaggt ggataaccga atccggattc tcattgagaa tggagtagct
     1381 gagcggcaaa gatctctctt tgttgtagtt ggggatcgag gaaaagatca ggtggtaata
     1441 cttcatcaca tgttatccaa agcaactgtg aaggctcggc cttcagtgct gtggtgttat
     1501 aagaaagagc tggggtttag cagtcaccgg aagaaaagaa tgcgacagct gcagaagaaa
     1561 ataaagaatg gaacactgaa cataaagcag gacgacccct ttgaactctt catagcagcc
     1621 acaaacattc gctactgcta ctacaacgag acccacaaga tcctgggcaa taccttcggc
     1681 atgtgtgtgc tgcaggattt tgaagcctta actccaaact tgctggccag gactgtagaa
     1741 acagtggaag gtggtgggct agtggtcatc ctcctacgga ccatgaactc actcaagcaa
     1801 ttgtacacag tgactatgga tgtgcattcc aggtacagaa ctgaggccca tcaggatgtg
     1861 gtgggaagat ttaatgaaag gtttattctg tctctggcct cttgtaagaa gtgtctcgtc
     1921 attgatgacc agctcaacat cctgcccatc tcctcccacg ttgccaccat ggaggccctg
     1981 cctccccaga ctccggatga gagtcttggt ccttctgatc tggagctgag ggagttgaag
     2041 gagagcttgc aggacaccca gcctgtgggt gtgttggtgg actgctgtaa gactctagac
     2101 caggccaaag ctgtcttgaa atttatcgag ggcatctctg aaaagaccct gaggagtact
     2161 gttgcactca cagctgctcg aggacgggga aaatctgcag ccctgggatt ggcgattgct
     2221 ggggcggtgg catttgggta ctccaatatc tttgttacct ccccaagccc tgataacctc
     2281 catactctgt ttgaatttgt atttaaagga tttgatgctc tgcaatatca ggaacatctg
     2341 gattatgaga ttatccagtc tctaaatcct gaatttaaca aagcagtgat cagagtgaat
     2401 gtatttcgag aacacaggca gactattcag tatatacatc ctgcagatgc tgtgaagctg
     2461 ggccaggctg aactagttgt gattgatgaa gctgccgcca tccccctccc cttggtgaag
     2521 agcctacttg gcccctacct tgttttcatg gcatccacca tcaatggcta tgagggcact
     2581 ggccggtcac tgtccctcaa gctaattcag cagctccgtc aacagagcgc ccagagccag
     2641 gtcagcacca ctgctgagaa taagaccacg acgacagcca gattggcatc agcgcggaca
     2701 ctgtatgagg tttccctcca ggagtcaatc cgatacgccc ctggggatgc agtggagaag
     2761 tggctgaatg acttgctgtg cctggattgc ctcaacatca ctcggatagt ctcaggctgc
     2821 cccttgcctg aagcttgtga actgtactat gttaatagag ataccctctt ttgctaccac
     2881 aaggcctctg aagttttcct ccaacggctt atggccctct acgtggcttc tcactacaag
     2941 aactctccca atgatctcca gatgctctcc gatgcacctg ctcaccatct cttctgcctt
     3001 ctgcctcccg tgccccccac ccagaatgcc cttccagaag tgcttgctgt tatccaggtg
     3061 tgccttgaag gggagatttc tcgccagtcc atcttgaaca gtctgtctcg aggcaagaag
     3121 gcttcagggg acctgattcc atggacagtg tcagaacagt tccaagatcc agactttggt
     3181 ggtctgtctg gtggaagggt cgttcgcatt gctgttcacc cagattatca agggatgggc
     3241 tatggcagcc gtgctctgca gctgctgcag atgtactatg aaggcaggtt tccttgtctg
     3301 gaggaaaagg tccttgagac accacaggaa attcacaccg taagcagcga ggctgtcagc
     3361 ttgttggaag aggtcatcac tccccggaag gacctgcctc ctttactcct caaattgaat
     3421 gagaggcctg ccgaacgcct ggattacctg ggtgtttcct atggcttgac ccccaggctc
     3481 ctcaagttct ggaaacgagc tggatttgtt cctgtttatc tgagacagac cccgaatgac
     3541 ctgaccggag agcactcgtg catcatgctg aagacgctca ctgatgagga tgaggctgac
     3601 cagggaggct ggcttgcagc cttctggaaa gatttccgac ggcggttcct agccttgctc
     3661 tcctaccagt tcagtacctt ctctccttcc ctggctctga acatcattca gaacaggaac
     3721 atggggaagc cagcccagcc tgccctgagc cgggaggagc tggaagcact cttcctcccc
     3781 tatgacctga agcggctgga gatgtattca cggaatatgg tggactatca cctcatcatg
     3841 gacatgatcc cggccatctc tcgcatctat ttcctgaacc agctggggga cctggccctg
     3901 tctgcggctc agtcggctct tctcttgggg attggcctgc agcataagtc tgtggaccag
     3961 ctggaaaagg agattgagct gccctcgggc cagttgatgg gacttttcaa ccggatcatc
     4021 cgcaaagttg tgaagctatt taatgaagtt caggaaaagg ccattgagga gcagatggtg
     4081 gcagcgaagg atgtggtcat ggagcccacg atgaagaccc tcagtgacga cctagatgaa
     4141 gcagcaaagg aatttcagga gaaacacaag aaggaagtag ggaagctgaa gagcatggac
     4201 ctctctgaat acataatccg tggggacgat gaagagtgga atgaagtttt gaacaaagct
     4261 gggccgaacg cctcgatcat cagcctgaaa agtgacaaga aaaggaagtt agaggccaaa
     4321 caagaaccca aacagagcaa gaagttgaag aacagagaga caaagaacaa aaaagatatg
     4381 aaactgaagc ggaagaaaga ctacaaggac gacgatgata agtgaccatg gtttaaaccc
     4441 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4501 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4561 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4621 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4681 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4741 gcgcattaaa taacttcgta taatgtatgc tatacgaagt tatatcgtat gcatgctttg
     4801 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     4861 cagaattcct tgaaatacat ccacaggtac acctccaatt gactcaaatg atgtcaatta
     4921 gtctatcaga agcttctaaa gccatgacat cattttctgg aattttccaa gctgtttaaa
     4981 ggcacagtca acttagtgta tgtaaacttc tgacccactg gaattgtgat acagtgaatt
     5041 ataagtgaaa taatctgtct gtaaacaatt gttggaaaaa tgacttgtgt catgcacaaa
     5101 gtagatgtcc taactgactt gccaaaacta ttgtttgtta acaagaaatt tgtggagtag
     5161 ttgaaaaacg agttttaatg actccaactt aagtgtatgt aaacttccga cttcaactgt
     5221 agggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5281 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5341 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5401 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     5461 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5521 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5581 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5641 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5701 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6361 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7201 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7261 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7321 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7381 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7441 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7501 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7561 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7621 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7681 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7741 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7801 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7861 gattacgcca agcttgcatg cctgcaggtc gactctagac ctatacagtt gaagtcggaa
     7921 gtttacatac acttaagttg gagtcattaa aactcgtttt tcaactactc cacaaatttc
     7981 ttgttaacaa acaatagttt tggcaagtca gttaggacat ctactttgtg catgacacaa
     8041 gtcatttttc caacaattgt ttacagacag attatttcac ttataattca ctgtatcaca
     8101 attccagtgg gtcagaagtt tacatacact aagttgactg tgcctttaaa cagcttggaa
     8161 aattccagaa aatgatgtca tggctttaga agcttctgat agactaattg acatcatttg
     8221 agtcaattgg aggtgtacct gtggatgtat ttcaagggga tccagacatg ataagataca
     8281 ttgatgagtt tggacaaacc acaactagaa tgcagcggcc gctagctcga gatcttaatt
     8341 aaactagtat cgatataact tcgtataatg tatgctatac gaagttatat gtacgggcca
     8401 gatatacgcg ttgacattga ttattgacta ggcttttgca aaaagctttg caaagatgga
     8461 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     8521 tgggaatt
//
