LOCUS       sequence-173651-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene for resistance to nourseothricin (Nat) inserted next to the 
            parS segments (INT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  AUTHORS   Saad H, Gallardo F, Dalvai M, Tanguy-le-Gac N, Lane D, Bystricky K
  TITLE     DNA dynamics during early double-strand break processing revealed by
            non-intrusive imaging of living cells.
  JOURNAL   PLoS Genet. 2014 Mar 13;10(3):e1004187. doi: 
            10.1371/journal.pgen.1004187. eCollection 2014 Mar.
  PUBMED    24625580
REFERENCE   2  (bases 1 to 4840)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Genet."; date: "2014-03-13"; volume: "10(3):e1004187. doi"; pages: "
            10.1371/journal.pgen.1004187. eCollection 2014 Mar"
COMMENT     Sequence Label: pANCH2-Nat
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             141..713
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      719..916
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1023..1042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1024..1042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1129..1146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1300..1888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1380..1399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2059..2850,2851..2919))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2682..2701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2920..3024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3092..3110
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3148..3170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3270..3289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3370..3388
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3370..3387
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     gene            join(4637..4840,1..916)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        join(4637..4840,1..140)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
ORIGIN
        1 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
       61 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
      121 acatccgaac ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca
      181 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
      241 ccgtcttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
      301 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
      361 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
      421 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
      481 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
      541 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
      601 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
      661 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
      721 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
      781 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
      841 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
      901 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagtgt cgaaaacgag
      961 ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt
     1021 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     1081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3061 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3121 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     3181 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     3241 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     3301 ccatatggac atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat
     3361 acacatacga tttaggtgac actatagaac gcggccgcca gctgaagctt ccggcgtcaa
     3421 cttctatcta ctccgaatcc caccattcca tgaatcgcgc gaccgacgtg tgcgttgtca
     3481 cgtgacaact ggcgctaaat atcggtgcgc cccgtactcg attggaggct caagggaatt
     3541 caatggtaga gcaccgatcc tcccggctca tcggcggatc ccgcgaagaa tcagccgaaa
     3601 cgtatcaaaa acaacaaatc acatatcaat caaaacaaat ctagccgcag ccgtatcaat
     3661 cttcaatgcc cgggcgttga aaactcatac gcccctattt cccgagggtg cagcatccgt
     3721 tttgcgtgaa acagcatgaa gaatatggcg caaaccattg cccgacaagg atttccgggc
     3781 gagtgcttat tttcgggaaa tgtttcacga tccgacccgt ttgggttgtt acatgcctct
     3841 gacgtatgga ttcagcgcca gtagcagttc atacctgcgg ctatctccgg catcaacagt
     3901 ccaccagcgc aggcaccggc tggcggccac gaagcaacct accctatcgg ccggcacgcg
     3961 tggaaacgca gggtcgcgct cctataaatg gggacatatc atgacagccc catgcagcct
     4021 ttgcgacaga gctcggcagc attgcgtccg cccttgaggg tccgcacgcc ggcaggaatt
     4081 tggccgcgca accgatcgtg ccaaggcctt cgcagcccca agccgtttct ttgcgacagc
     4141 aacggggcgc tcctcgccat agagtcgcca tcgcttgaac ttcccggcat cgcgtcctgt
     4201 tgtcacgtga caactcaccg atggacacac ggacaaacca cctcggcgaa aactatcaac
     4261 ttcgtgcgtt tccccgacaa tatgcctcca cgccgcgagc gtcgaccagc aacaactatt
     4321 cttaactgac cacaaatccc ctatccgaga cagcaccttg ggattgtttg tcacgtgaca
     4381 actcatcttc ccaggtagac gtccgcgatt tagcgcccgt gtatcgataa ttagagtgaa
     4441 gcgattaccc cgcgctccag gtcgcaatcg aaaactttcg ctcgacgtag cactcggagg
     4501 aatcgatatc cggcgacttg tcacgtgaca accggctctt cctcgttacc ggggtctcag
     4561 cgccccggat ttgggttaat taaggcgcgc cagatctgtt tagcttgcct cgtccccgcc
     4621 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     4681 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     4741 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     4801 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
//
