LOCUS       sequence-173633-        5296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gene for resistance to hygromycin (Hyg) inserted next to the parS 
            segments (INT). Also known as pANCH2-Hph.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5296)
  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 5296)
  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: pFG4
FEATURES             Location/Qualifiers
     source          1..5296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..77
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     311..328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     415..434
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        415..433
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            complement(541..2116)
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     terminator      complement(541..738)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(744..1772)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             complement(744..1772)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(1773..2116)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        complement(3365..3383)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3366..3383)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3464..3483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3583..3605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3643..3661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3834..3902,3903..4694)
                     /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     complement(4052..4071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4865..5296,1..157)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
       61 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      181 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      241 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      301 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      361 tctccccgcg cgttggccga ttcattaatg caggttaacc tggcttatcg aaattaatac
      421 gactcactat agggagaccg gcagatccgc ggccgcatag gccactagtg gatctgatat
      481 catcgatgaa ttcgagctcg ttttcgacac tggatggcgg cgttagtatc gaatcgacag
      541 cagtatagcg accagcattc acatacgatt gacgcatgat attactttct gcgcacttaa
      601 cttcgcatct gggcagatga tgtcgaggcg aaaaaaaata taaatcacgc taacatttga
      661 ttaaaataga acaactacaa tataaaaaaa ctatacaaat gacaagttct tgaaaacaag
      721 aatcttttta ttgtcagtac tgattattcc tttgccctcg gacgagtgct ggggcgtcgg
      781 tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg
      841 gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc
      901 tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga
      961 ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc
     1021 tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg
     1081 gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg
     1141 cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact
     1201 tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg
     1261 acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg
     1321 aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc
     1381 gtctggctaa gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca
     1441 gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg
     1501 caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg
     1561 gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt
     1621 acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc
     1681 tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca
     1741 gacgtcgcgg tgagttcagg ctttttaccc atggttgttt atgttcggat gtgatgtgag
     1801 aactgtatcc tagcaagatt ttaaaaggaa gtatatgaaa gaagaacctc agtggcaaat
     1861 cctaaccttt tatatttctc tacaggggcg cggcgtgggg acaattcaac gcgtctgtga
     1921 ggggagcgtt tccctgctcg caggtccgca gcgaggagcc gtaatttttg cttcgcgccg
     1981 tgcggccatc aaaatgtatg gatgcaaatg attatacatg gggatgtatg ggctaaatgt
     2041 acgggcgaca gtcacatcat gcccctgagc tgcgcacgtc aagactgtca aggagggtat
     2101 tctgggcctc catgtcgctg gccgggtgac ccggcgggga cgaggcaagc taaacagatc
     2161 tggcgcgcct taattaaccc aaatccgggg cgctgagacc ccggtaacga ggaagagccg
     2221 gttgtcacgt gacaagtcgc cggatatcga ttcctccgag tgctacgtcg agcgaaagtt
     2281 ttcgattgcg acctggagcg cggggtaatc gcttcactct aattatcgat acacgggcgc
     2341 taaatcgcgg acgtctacct gggaagatga gttgtcacgt gacaaacaat cccaaggtgc
     2401 tgtctcggat aggggatttg tggtcagtta agaatagttg ttgctggtcg acgctcgcgg
     2461 cgtggaggca tattgtcggg gaaacgcacg aagttgatag ttttcgccga ggtggtttgt
     2521 ccgtgtgtcc atcggtgagt tgtcacgtga caacaggacg cgatgccggg aagttcaagc
     2581 gatggcgact ctatggcgag gagcgccccg ttgctgtcgc aaagaaacgg cttggggctg
     2641 cgaaggcctt ggcacgatcg gttgcgcggc caaattcctg ccggcgtgcg gaccctcaag
     2701 ggcggacgca atgctgccga gctctgtcgc aaaggctgca tggggctgtc atgatatgtc
     2761 cccatttata ggagcgcgac cctgcgtttc cacgcgtgcc ggccgatagg gtaggttgct
     2821 tcgtggccgc cagccggtgc ctgcgctggt ggactgttga tgccggagat agccgcaggt
     2881 atgaactgct actggcgctg aatccatacg tcagaggcat gtaacaaccc aaacgggtcg
     2941 gatcgtgaaa catttcccga aaataagcac tcgcccggaa atccttgtcg ggcaatggtt
     3001 tgcgccatat tcttcatgct gtttcacgca aaacggatgc tgcaccctcg ggaaataggg
     3061 gcgtatgagt tttcaacgcc cgggcattga agattgatac ggctgcggct agatttgttt
     3121 tgattgatat gtgatttgtt gtttttgata cgtttcggct gattcttcgc gggatccgcc
     3181 gatgagccgg gaggatcggt gctctaccat tgaattccct tgagcctcca atcgagtacg
     3241 gggcgcaccg atatttagcg ccagttgtca cgtgacaacg cacacgtcgg tcgcgcgatt
     3301 catggaatgg tgggattcgg agtagataga agttgacgcc ggaagcttca gctggcggcc
     3361 gcgttctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt
     3421 agccgcgttc taacgacaat atgtccatat ggtgcactct cagtacaatc tgctctgatg
     3481 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3541 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3601 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3661 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3961 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4141 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4201 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4441 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4501 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4561 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4681 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4741 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4801 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4861 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4921 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4981 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5041 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5101 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5161 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5221 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5281 aagggagaaa ggcgga
//
