LOCUS       sequence-7289-ex        8703 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 8703)
  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 8703)
  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: pFS177
FEATURES             Location/Qualifiers
     source          1..8703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1753..2534
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(2617..2633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2617..2633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2630..2652)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2641..2657
                     /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)."
     promoter        complement(join(2665..2671,2672..2689,2690..2695))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2710..2731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2848..2865)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3019..3607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3099..3118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3778..4569,4570..4638))
                     /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     4401..4420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4639..4743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4811..4829
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4867..4889)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4989..5008
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5025..5480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5162..5183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5374..5393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5678..5700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5692..5709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5693..5709
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5960..6367
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6368..7462
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 tgcaggtcga ctctagagga tctcatatga caaccagtca ccagcctcag gacagataca
       61 aagctgtctg gcttatcttc ttcatgctgg gtctgggaac gctgctcccg tggaattttt
      121 tcatgacggc cactcagtat ttcacaaacc gcctggacat gtcccagaat gtgtccttgg
      181 tcactgctga actgagcaag gacgcccagg cgtcagccgc ccctgcagca cccttgcctg
      241 agcggaactc tctcagtgcc atcttcaaca atgtcatgac cctatgtgcc atgctgcccc
      301 tgctgttatt cacctacctc aactccttcc tgcatcagag gatcccccag tccgtacgga
      361 tcctgggcag cctggtggcc atcctgctgg tgtttctgat cactgccatc ctggtgaagg
      421 tgcagctgga tgctctgccc ttctttgtca tcaccatgat caagatcgtg ctcattaatt
      481 catttggtgc catcctgcag ggcagcctgt ttggtctggc tggccttctg cctgccagct
      541 acacggcccc catcatgagt ggccagggcc tagcaggctt ctttgcctcc gtggccatga
      601 tctgcgctat tgccagtggc tcggaactat cagaaagtgc cttcggctac tttatcacag
      661 cctgtgctgt tatcattttg accatcatct gttacctggg cctgccccgc ctggaattct
      721 accgctacta ccagcagctc aagcttgaag gacccgggga gcaggagacc aagttggacc
      781 tcattagcaa aggagaggag ccaagagcag gcaaagagga atctggagtt tcagtctcca
      841 actctcagcc caccaatgaa agccactcta tcaaagccat cctgaaaaat atctcagtcc
      901 tggctttctc tgtctgcttc atcttcacta tcaccattgg gatgtttcca gccgtgactg
      961 ttgaggtcaa gtccagcatc gcaggcagca gcacctggga acgttacttc attcctgtgt
     1021 cctgtttctt gactttcaat atctttgact ggttgggccg gagcctcaca gctgtattca
     1081 tgtggcctgg gaaggacagc cgctggctgc caagcctggt gctggcccgg ctggtgtttg
     1141 tgccactgct gctgctgtgc aacattaagc cccgccgcta cctgactgtg gtcttcgagc
     1201 acgatgcctg gttcatcttc ttcatggctg cctttgcctt ctccaacggc tacctcgcca
     1261 gcctctgcat gtgcttcggg cccaagaaag tgaagccagc tgaggcagag accgcaggag
     1321 ccatcatggc cttcttcctg tgtctgggtc tggcactggg ggctgttttc tccttcctgt
     1381 tccgggcaat tgtgtgagag ctcgaattcc aaccttccaa ttcattaaat cgagtcctcc
     1441 atcagatgat ttacaaggtt ttgcatagaa tccctaagta agagaagttg ttagtgattt
     1501 cttcaagttt actaaaattt acatacaaat ggatgatctc ttctccattg tttcctttct
     1561 tcttgcaaac gcgttttaca aagagatgac atgctatttt ctgattattt ttttctatag
     1621 ttttctattt tgtacgtcct tgggagtccg aaatgtaaaa tcggggtatg gttagtagcg
     1681 gcagcgttca ttatggtata taaaagataa aaagtagggg caagcgagac agaatgggat
     1741 acaagggcat cgtctataat tatagctaaa aattgtattt taatttgtat tttttgtaat
     1801 tttatttctc attgttttac ttaaaatgaa tgcgaattag agaaaacttc aacgaaatgt
     1861 caaaataagc tcagcaaaat atacaatttt agggaaagcg atcagcaact tattctccgt
     1921 tgtagattct ttttcttcat aaattccaac aatgtaactg ccaattgatt tttctctaga
     1981 tgctaataaa ttgaaactaa tgaatataat aacacaggtt ttgataaatt gtaaactctt
     2041 actcaaaaaa agaatgttaa aactaattta acctttttat aaaggctcat ggcgttaaag
     2101 aggatagtaa aaattctaca attttttaat ttaatgtcac aattgtcttg cgaatataat
     2161 tctattaatt ttcgctaact gctgttttag tactattact ttcaatatgc cttatatgca
     2221 actttaattc atgaagaatg caatattttt gcaatctttc ttgtactgag cgctttttat
     2281 catttagtac tcattattta atttttcagt aaagggcaat aaataaattt gtagaagatg
     2341 caatgtaatc tcctctatcc ttttgctcat atgtttatga gtatacctag tctagaaagg
     2401 cttgtattta aaatattatt caataaaaat tcacaatttt tacgacatgt gctaccattc
     2461 acttaacttc ctgattataa aattggttcg tttatactaa ttacttaagt acctttaact
     2521 aaacaaaatg cctatatata tattaattta caatgagtgt cagataagtc actatgtccg
     2581 agtggttaag gagttagact cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     2641 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2701 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2761 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2821 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2881 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2941 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3001 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3061 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3421 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4741 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4801 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4861 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4921 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4981 taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa
     5041 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5101 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt
     5161 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5221 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
     5281 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5341 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5401 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5461 tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac
     5521 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5581 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5641 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5701 acggccagtg ccaagcttgt cgaggagaac ttctagtata tctacatacc taatattatt
     5761 gccttattaa aaatggaatc ccaacaatta catcaaaatc cacattctct tcaaaatcaa
     5821 ttgtcctgta cttccttgtt catgtgtgtt caaaaacgtt atatttatag gataattata
     5881 ctctatttct caacaagtaa ttggttgttt ggccgagcgg tctaaggcgc ctgattcaag
     5941 aaatatcttg accgcagtta actgtgggaa tactcaggta tcgtaagatg caagagttcg
     6001 aatctcttag caaccattat ttttttcctc aacataacga gaacacacag gggcgctatc
     6061 gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc gcctgacgca
     6121 tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagccg cggaaccggc
     6181 ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat acttgaagtt
     6241 gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt
     6301 tctaactttt cttacctttt acatttcagc aatatatata tatatatttc aaggatatac
     6361 cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga
     6421 aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa
     6481 gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgtcccact
     6541 tccagatgag gcgctggaag cctccaagaa ggttgatgcc gttttgttag gtgctgtggg
     6601 tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa
     6661 agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga
     6721 cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt
     6781 agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga
     6841 tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc
     6901 cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc
     6961 ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgaattcc ctacattgaa
     7021 ggttcaacat caattgattg attctgccgc catgatccta gttaagaacc caacccacct
     7081 aaatggtatt ataatcacca gcaacatgtt tggtgatatc atctccgatg aagcctccgt
     7141 tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa
     7201 caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa
     7261 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt
     7321 gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag
     7381 aactggtgat ttaggtggtt ccaacagtac cacggaagtc ggtgatgctg tcgccgaaga
     7441 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     7501 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     7561 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     7621 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     7681 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga
     7741 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa
     7801 atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct
     7861 tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg
     7921 gccttaacga cgtagtcgat cgacaagctt gcatgcccta caacaactaa gaaaatggct
     7981 atcatgcgga agctggtgag aagaacagca tcgggacaag ggaaggaaga acaaagacaa
     8041 agaaaacaaa agaaagcaat tgaaaacaaa acaaaacaat tttcattcct tctcttatca
     8101 ttccttttct tttcttttct ctcattcaac gcactccatc gtatccgtat tcctcttatt
     8161 ttttctcttt ctctatatcc atttctttct ctctaggtgt gtcctctctc tctcttcaat
     8221 ttctctactc cgcattccaa cgcatccttc ccccaacctc ccatttcctc cttacggccc
     8281 gatagcgatc gtctttccct cgctatcact cgctaccggc ccctcctctg caccgtaacc
     8341 tcctacgtat ttaccatatc ataaagtttt ttccgacgct tatcgctgac cccctgtcgc
     8401 cctcctattg gcttccggat tatcttcttg tccataaggt gatccatgct tcctgaagat
     8461 tcccgaaatg tgtccacttt ggcggggaat cattccatcc acttctttct ctctcgcttt
     8521 cctcattcgg cgctcccctt ccgcgtctca ttggtcttcc gctccgtttt tgctttgccg
     8581 atgttacttg gggagaggtg cgataatcct ttcgcaaaaa ctcggtttga cgcctcccat
     8641 ggtataaata gtgggtggtg gacaggtgcc ttcgcttttc ttttctttaa gcaagagaat
     8701 tcc
//
