LOCUS       sequence-173625-        4708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene for resistance to nourseothricin (Nat) inserted next to the 
            parS segments (INT). Also known as pANCH1-Nat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4708)
  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 4708)
  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: pFG2
FEATURES             Location/Qualifiers
     source          1..4708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            521..1640
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        521..864
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             865..1437
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1443..1640
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1747..1766)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1748..1766)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1853..1870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2024..2612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2104..2123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2783..3574,3575..3643))
                     /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     3406..3425
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3644..3748)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3816..3834
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3872..3894)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3994..4013
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4094..4112
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4094..4111
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgctgtacc tcgcccggca atcgcgcgct ttatgcgcat aaacctagcc ggcgtgttcc
       61 gcaatcgcat acccctttcg cgacccgaga cagccctcac gagacagccc tcacgagaca
      121 gccctcacga gacagccctc acgagacatc ggctgcgccg gtcgtcggcc ccgcgaagcc
      181 acccgcccga ctttatgcgc ataaacaacc actcttcgac gcgtcgccga acacaggaca
      241 ctcccctctc gcgatcactg cgcatcgcca aaccgaaacc gcctcagcga cctatcaaga
      301 ggaaccccac accgatctcg attgcccccc atcaggcccc tcgatgcaga ctcgctacaa
      361 tacaggcctc acccctcctg ccttgaacga tgatcacgat ctaccacaac ccccgatgct
      421 cgaaatcccg cgagacgctt gcgttggtcg agtcgctgaa taccgccggc gcgccagatc
      481 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
      541 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      601 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      661 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
      721 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
      781 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
      841 catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc ttaccggtac
      901 cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc
      961 gacaccgtct tccgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg
     1021 gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg
     1081 gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg
     1141 ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag
     1201 gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag
     1261 ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg
     1321 gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac
     1381 gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca
     1441 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     1501 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     1561 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     1621 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa
     1681 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     1741 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     1801 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1861 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1921 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1981 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2041 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2101 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2161 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2221 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2281 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2341 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2401 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2461 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2521 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2581 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2641 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2701 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2761 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2821 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2881 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2941 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3001 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3061 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3121 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3181 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3241 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3301 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3361 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3421 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3481 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3541 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3601 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3661 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3721 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3781 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     3841 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     3901 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     3961 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4021 tgcaccatat ggacatattg tcgttagaac gcggctacaa ttaatacata accttatgta
     4081 tcatacacat acgatttagg tgacactata gaacgcggcc gccagctgaa gcttcgtacg
     4141 ctgcaggtcg acggatcccc ccattcggac gatcggttta tgcgcataaa cgagcggtgg
     4201 atgttcgatg gctcgtttgg acggtgtggt gaagaggaag cggctgatgc cgagtgatcg
     4261 atgccctgct caggattacg tcttcatggg cgtatcgtcg gggccgttcg tggcagagat
     4321 atctcgacgc tctctccttt cgcgggacga gccgcgttgc ctgcgagccg cgagtttgtg
     4381 cgcataaaga ctgcggaggc cgatcgatgg gcttcgtcgc tagagcgcat ggtattcact
     4441 cgcgagcgtc cggcgtgagc tgcgtgtcgt agtcaggtgc gtagatcacg aggtgcgtcg
     4501 ttgaatggac gtgcatggag gcaatgtcct gccgggccgt agtgtttatg cgcataaacc
     4561 agaggcaacg ataggccgcg acgaagcgcc gttgacagag actacgcctg agtcaggcgg
     4621 gcaatgacgt catgtcgggg acgacggtct atcgatatag ccatcgcgca gtgccgttgc
     4681 gtgtcgacat atgtccatgc cacgagcc
//
