LOCUS       sequence-7290-ex        5925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5925)
  AUTHORS   Sivakumar S, Porter-Goff M, Patel PK, Benoit K, Rhind N
  TITLE     In vivo labeling of fission yeast DNA with thymidine and thymidine 
            analogs.
  JOURNAL   Methods. 2004 Jul . 33(3):213-9.
  PUBMED    15157888
REFERENCE   2  (bases 1 to 5925)
  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: "Methods. 
            2004 Jul . 33(3):213-9."
COMMENT     Sequence Label: pFS255
FEATURES             Location/Qualifiers
     source          1..5925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(143..1273)
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMIQTHVTTPGSIPTICDLARTFAREMGEAN"
     gene            2102..3458
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        2102..2445
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2446..3255
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2513..2532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      3261..3458
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3562..3581)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3563..3581)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3668..3685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3839..4427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3919..3938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4598..5389,5390..5458))
                     /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     5221..5240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5459..5563)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5631..5649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5687..5709)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5809..5828
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(5909..5925,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(5909..5925,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta
       61 aggcgcgcca gatcgatctt cgaatgcatc gcgcgcaccg tacgtctcga ggaattcctg
      121 caggatccgg ggatcccgtg tttcagttag cctcccccat ctcccgggca aacgtgcgcg
      181 ccaggtcgca gatcgtcggt atggagcctg gggtggtgac gtgggtctgg atcatcccgg
      241 aggtaagttg cagcagggcg tcccggcagc cggcgggcga ttggtcgtaa tccaggataa
      301 agacgtgcat gggacggagg cgtttggcca agacgtccaa ggcccaggca aacacgttgt
      361 acaggtcgcc gttgggggcc agcaactcgg gggcccgaaa cagggtaaat aacgtgtccc
      421 cgatatgggg tcgtgggccc gcgttgctct ggggctcggc accctggggc ggcacggccg
      481 tccccgaaag ctgtccccaa tcctcccgcc acgacccgcc gccctgcaga taccgcaccg
      541 tattggcaag cagcccgtaa acgcggcgaa tcgcggccag catagccagg tcaagccgct
      601 cgccggggcg ctggcgtttg gccaggcggt cgatgtgtct gtcctccgga agggccccca
      661 acacgatgtt tgtgccgggc aaggtcggcg ggatgagggc cacgaacgcc agcacggcct
      721 ggggggtcat gctgcccata aggtatcgcg cggccgggta gcacaggagg gcggcgatgg
      781 gatggcggtc gaagatgagg gtgagggccg ggggcggggc atgtgaactc ccagcctccc
      841 ccccgatatg aggagccaga acggcgtcgg tcacggcata aggcatgccc attgttatct
      901 gggcgcttgt cattaccacc gccgcgtccc cggccgatat ctcaccctgg tcgaggcggt
      961 gttgtgtggt gtagatgttc gcgattgtct cggaagcccc cagcacccgc cagtaagtca
     1021 tcggctcggg tacgtagacg atatcgtcgc gcgaacccag ggccaccagc agttgcgtgg
     1081 tggtggtttt ccccatcccg tggggaccgt ctatataaac ccgcagtagc gtgggcattt
     1141 tctgctccgg gcggacttcc gtggcttctt gctgccggcg agggcgcaac gccgtacgtc
     1201 ggttgctatg gccgcgagaa cgcgcagcct ggtcgaacgc agacgcgtgc tgatggccgg
     1261 ggtacgaagc catacgggat cctctagagt cgacctgcag caattctctt gcttaaagaa
     1321 aagaaaagcg aaggcacctg tccaccaccc actatttata ccatgggagg cgtcaaaccg
     1381 agtttttgcg aaaggattat cgcacctctc cccaagtaac atcggcaaag caaaaacgga
     1441 gcggaagacc aatgagacgc ggaaggggag cgccgaatga ggaaagcgag agagaaagaa
     1501 gtggatggaa tgattccccg ccaaagtgga cacatttcgg gaatcttcag gaagcatgga
     1561 tcaccttatg gacaagaaga taatccggaa gccaatagga gggcgacagg gggtcagcga
     1621 taagcgtcgg aaaaaacttt atgatatggt aaatacgtag gaggttacgg tgcagaggag
     1681 gggccggtag cgagtgatag cgagggaaag acgatcgcta tcgggccgta aggaggaaat
     1741 gggaggttgg gggaaggatg cgttggaatg cggagtagag aaattgaaga gagagagagg
     1801 acacacctag agagaaagaa atggatatag agaaagagaa aaaataagag gaatacggat
     1861 acgatggagt gcgttgaatg agagaaaaga aaagaaaagg aatgataaga gaaggaatga
     1921 aaattgtttt gttttgtttt caattgcttt cttttgtttt ctttgtcttt gttcttcctt
     1981 cccttgtccc gatgctgttc ttctcaccag cttccgcatg atagccattt tcttagttgt
     2041 tgtagggcat gcaagctgat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag
     2101 cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg
     2161 atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat
     2221 acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga
     2281 gcagggaaac gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag
     2341 aaatataaaa ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat
     2401 cttgctagga tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag
     2461 actcacgttt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2521 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc
     2581 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2641 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2701 atccgtactc ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc
     2761 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2821 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2881 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2941 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca
     3001 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     3061 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     3121 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     3181 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     3241 gatgagtttt tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     3301 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     3361 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     3421 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     3481 gccatccagt ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg
     3541 cggccgcgga tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta
     3601 acctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3661 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3721 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3781 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3841 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3901 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3961 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4021 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4081 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4141 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4201 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4261 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4321 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5581 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5641 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     5701 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     5761 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     5821 agattgtact gagagtgcac catatggaca tattgtcgtt agaacgcggc tacaattaat
     5881 acataacctt atgtatcata cacatacgat ttaggtgaca ctata
//
