LOCUS       sequence-7287-ex        7282 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 7282)
  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 7282)
  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: pFS118
FEATURES             Location/Qualifiers
     source          1..7282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(143..211,212..1003)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(361..380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1045..1554)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1240..1261)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1448..1467
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1665..2253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2154..2173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2314..2335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2350..2355,2356..2373,2374..2380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2388..2404
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2393..2415
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2412..2428
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2412..2428
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2539..3669)
                     /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"
     CDS             4982..5776
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      6247..7028
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(7083..7100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7083..7099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7092..7114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gttaactacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
       61 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      121 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      181 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      241 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      301 gatccttgag agttttcgcc ccgaagaacg ttctccaatg atgagcactt ttaaagttct
      361 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
      421 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      481 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      541 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      601 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      661 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      721 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      781 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      841 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      901 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      961 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1021 ctcatatata ctttagattg atttaccccg gttgataatc agaaaagccc caaaaacagg
     1081 aagattgtat aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta
     1141 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1201 aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1261 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1321 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta
     1381 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcg aacgtggcga
     1441 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     1501 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg
     1561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1801 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2281 ttcctggcct tttgctggcc ttttgctcac atgtaatgtg agttagctca ctcattaggc
     2341 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2401 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctacgta atacgactca
     2461 ctagtgggca gatcttcgaa tgcatcgcgc gcaccgtacg tctcgaggaa ttcctgcagg
     2521 atccggggat cccgtgtttc agttagcctc ccccatctcc cgggcaaacg tgcgcgccag
     2581 gtcgcagatc gtcggtatgg agcctggggt ggtgacgtgg gtctggatca tcccggaggt
     2641 aagttgcagc agggcgtccc ggcagccggc gggcgattgg tcgtaatcca ggataaagac
     2701 gtgcatggga cggaggcgtt tggccaagac gtccaaggcc caggcaaaca cgttgtacag
     2761 gtcgccgttg ggggccagca actcgggggc ccgaaacagg gtaaataacg tgtccccgat
     2821 atggggtcgt gggcccgcgt tgctctgggg ctcggcaccc tggggcggca cggccgtccc
     2881 cgaaagctgt ccccaatcct cccgccacga cccgccgccc tgcagatacc gcaccgtatt
     2941 ggcaagcagc ccgtaaacgc ggcgaatcgc ggccagcata gccaggtcaa gccgctcgcc
     3001 ggggcgctgg cgtttggcca ggcggtcgat gtgtctgtcc tccggaaggg cccccaacac
     3061 gatgtttgtg ccgggcaagg tcggcgggat gagggccacg aacgccagca cggcctgggg
     3121 ggtcatgctg cccataaggt atcgcgcggc cgggtagcac aggagggcgg cgatgggatg
     3181 gcggtcgaag atgagggtga gggccggggg cggggcatgt gaactcccag cctccccccc
     3241 gatatgagga gccagaacgg cgtcggtcac ggcataaggc atgcccattg ttatctgggc
     3301 gcttgtcatt accaccgccg cgtccccggc cgatatctca ccctggtcga ggcggtgttg
     3361 tgtggtgtag atgttcgcga ttgtctcgga agcccccagc acccgccagt aagtcatcgg
     3421 ctcgggtacg tagacgatat cgtcgcgcga acccagggcc accagcagtt gcgtggtggt
     3481 ggttttcccc atcccgtggg gaccgtctat ataaacccgc agtagcgtgg gcattttctg
     3541 ctccgggcgg acttccgtgg cttcttgctg ccggcgaggg cgcaacgccg tacgtcggtt
     3601 gctatggccg cgagaacgcg cagcctggtc gaacgcagac gcgtgctgat ggccggggta
     3661 cgaagccata cgggatcctc tagagtcgac ctgcagcaat tctcttgctt aaagaaaaga
     3721 aaagcgaagg cacctgtcca ccacccacta tttataccat gggaggcgtc aaaccgagtt
     3781 tttgcgaaag gattatcgca cctctcccca agtaacatcg gcaaagcaaa aacggagcgg
     3841 aagaccaatg agacgcggaa ggggagcgcc gaatgaggaa agcgagagag aaagaagtgg
     3901 atggaatgat tccccgccaa agtggacaca tttcgggaat cttcaggaag catggatcac
     3961 cttatggaca agaagataat ccggaagcca ataggagggc gacagggggt cagcgataag
     4021 cgtcggaaaa aactttatga tatggtaaat acgtaggagg ttacggtgca gaggaggggc
     4081 cggtagcgag tgatagcgag ggaaagacga tcgctatcgg gccgtaagga ggaaatggga
     4141 ggttggggga aggatgcgtt ggaatgcgga gtagagaaat tgaagagaga gagaggacac
     4201 acctagagag aaagaaatgg atatagagaa agagaaaaaa taagaggaat acggatacga
     4261 tggagtgcgt tgaatgagag aaaagaaaag aaaaggaatg ataagagaag gaatgaaaat
     4321 tgttttgttt tgttttcaat tgctttcttt tgttttcttt gtctttgttc ttccttccct
     4381 tgtcccgatg ctgttcttct caccagcttc cgcatgatag ccattttctt agttgttgta
     4441 gggcatgcaa gcttagctac aaatcccact ggctatatgt atgcatttgt gttaaaaaag
     4501 tttgtataga ttatttaatc tactcagcat tctttctcta aataggaatt tgttacttaa
     4561 tggagaaaaa aatgtttcga tttacctagt gtatttgttt gtatactcac gtttaatttc
     4621 aaacatccat tctatcttgt gtaatttttg gcatggtgaa aaagataatc agccttataa
     4681 tctttacaaa agtaagaaat tctgtaaata agccttaatg cccttgcttt aaattaaaat
     4741 ggttcttttt catgataatg tttgcacttt gtgaatatat tttagatagt tctgtgaggt
     4801 ataattaaga tgttttagag acttatacaa ttttgtcttt ataaattctt aattgatttt
     4861 accatcccag tttaactatg cttcgtcggc atctctgcac atgtcgtgtt ttcttaccgt
     4921 attgtcctac caagaacctc ttttttgctt ggatcgaaat taaaggttta aaagcaaagt
     4981 tatggatgct agagtatttc aaagctattc agctagagct gaggggatga aaaatcccat
     5041 tgccaaggaa ttgttggctt tgatggaaga aaagcaaagc aacttgtcag tcgcggtcga
     5101 tttgacgaag aaatccgaaa tcttagaatt ggtagataaa attggaccct atgtctgtgt
     5161 tatcaagaca catattgacg ttgtcgagga tttcgaccag gatatggtag aaaaactggt
     5221 ggccttaggt aaaaagcatc gttttcttat ctttgaggat cgcaaattcg cagacattgg
     5281 aaataccgtc aagctacaat atgcatctgg tgtgtacaaa attgcttctt gggctcatat
     5341 cacaaattgc catacagtgc caggcgaggg tattatacaa ggcctcaaag aagttggttt
     5401 acctttggga cgtggtctct tgcttttggc tgaaatgtct tccaaaggct ctttggctac
     5461 tggttcctac acagagaaaa ccttagaatg gtttgagaag cataccgatt tttgctttgg
     5521 ctttatagct ggtcgtcgat ttcctaacct tcaaagcgac tacataacta tgtcccctgg
     5581 tatcggcttg gatgttaaag gagacgggct gggacagcaa tatcgtactc ctgaagaagt
     5641 gattgtaaac tgcggtagcg atatcatcat tgttggtcgt ggagtctatg gagctggtcg
     5701 taatcctgtt gtcgaagcca agagatatag agaagctggt tggaaggcat atcagcaaag
     5761 actttctcag cattaaaaaa agactaatgt aaaatttttt tggttggtta ttgaaaaagt
     5821 cgatgccttg tttgcgtttg ttttcctagg cgttttatgt cagaaggcat ttagaattag
     5881 tatacaagta ctctttggta aaattttatg tagcgactaa aatattaact attatagata
     5941 aacaccttgg gaataaaaag taatttgcta tagtaattta ttaaacatgc tcctacaaca
     6001 ttaccacaat cttttctctt ggattgacat tgaataagaa aagagtgaat ttttttagac
     6061 ttgtaatgat aactatgtac aaagccaatg aaagatgtat gtagatgaat gtaaaatacc
     6121 atgtagacaa acaagataaa acttggttat aaacattggt gttggaacag aataaattag
     6181 atgtcaaaaa gtttcgtcaa tatcacaagc ttcccatggt gacgtcaccg gttctagata
     6241 cctaggtagg cattttgttt agttaaaggt acttaagtaa ttagtataaa cgaaccaatt
     6301 ttataatcag gaagttaagt gaatggtagc acatgtcgta aaaattgtga atttttattg
     6361 aataatattt taaatacaag cctttctaga ctaggtatac tcataaacat atgagcaaaa
     6421 ggatagagga gattacattg catcttctac aaatttattt attgcccttt actgaaaaat
     6481 taaataatga gtactaaatg ataaaaagcg ctcagtacaa gaaagattgc aaaaatattg
     6541 cattcttcat gaattaaagt tgcatataag gcatattgaa agtaatagta ctaaaacagc
     6601 agttagcgaa aattaataga attatattcg caagacaatt gtgacattaa attaaaaaat
     6661 tgtagaattt ttactatcct ctttaacgcc atgagccttt ataaaaaggt taaattagtt
     6721 ttaacattct ttttttgagt aagagtttac aatttatcaa aacctgtgtt attatattca
     6781 ttagtttcaa tttattagca tctagagaaa aatcaattgg cagttacatt gttggaattt
     6841 atgaagaaaa agaatctaca acggagaata agttgctgat cgctttccct aaaattgtat
     6901 attttgctga gcttattttg acatttcgtt gaagttttct ctaattcgca ttcattttaa
     6961 gtaaaacaat gagaaataaa attacaaaaa atacaaatta aaatacaatt tttagctata
     7021 attatagacc taggtgagct ctggtaccct ctagtcaagg ccttaagtga gtcgtattac
     7081 ggactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     7141 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     7201 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcttcgcttg gtaataaagc
     7261 ccgcttcggc gggctttttt tt
//
