LOCUS       sequence-7350-ex        6660 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 6660)
  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 6660)
  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: pFS181
FEATURES             Location/Qualifiers
     source          1..6660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4400..4420)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4400..4418)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4439..4455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4439..4455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4452..4474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4463..4479
                     /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(4487..4493,4494..4511,4512..4517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4670..4687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4841..5429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5600..6391,6392..6460))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6223..6242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6461..6565)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6633..6651
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatacgat atcccaatct gtagtataaa aagtgcaaca gtgttgagtt tcccgaaacc
      241 aggcattttt gcggtagggt ctgaaacttt cacgaaacca tttcacaaca aaacttcttc
      301 aacaacatgt gtgcaaagaa aatcgtcgtc ttaccaggag accatattgg ccctgaaatt
      361 gttgcttctg ccttggaggt tttgaaagtc gttgagaaga agcgacctga gttaaaactc
      421 gagtttgaag aacacaagat tggaggtgcc tctattgatg cctatggaac ccctttgact
      481 gacgagactg tgaaggcttg tttggaagct gacggtgttc ttttgggtgc cgttggtggt
      541 cctgaatgga ccaaccccaa ttgtcgtcct gagcaaggtt tattgaagct tcgtaagagt
      601 atgggtgttt gggccaacct tcgaccttgc aactttgcca gcaagtcttt agtcaagtac
      661 agccctttga agcctgaaat cgttgaaggt gtcgattttt gtgttgtacg agaacttact
      721 ggaggttgtt actttggtga gcgcactgag gacaacggat cgggttatgc tatggacact
      781 tggccttaca gtttggaaga agtttctcgt attgctcgtt tggctgcttg gttagctgaa
      841 acttccaacc ctcctgctcc cgtcacatta ctcgacaaag ctaatgtttt ggcaacttct
      901 cgtctttggc gcaagactgt agctaagatt tttaaggagg aatatcctca ccttacctta
      961 aaaaaccagc tcattgactc cgcagccatg cttttggtca agagccctcg tacacttaac
     1021 ggtgttgttt tgactgacaa cttgtttggt gacattattt cagatgaggc ttctgtcatt
     1081 cctggtagct tgggcctttt gccttctgcc tccctttccg gtgtggtagg aaaatcagaa
     1141 gaaaaggttc attgtttggt tgagcccatt cacggtagcg ctcccgatat cgctggtaag
     1201 ggcattgtta atcctgttgg tacaatttta tctgcttccc ttctccttcg ttatggtttg
     1261 aatgctccta aggaggctga agctatcgaa gccgccgtac gcaaggtctt ggatgatact
     1321 tcaattggtg gacgtggtct ttatactcgc gatttgggag gtgaggcttc taccgctgat
     1381 attactaagg ctgttgttga agaacttgaa aaaattttgt aatagggttt aatgtagaat
     1441 aaattcatat gatttgattt gacctaattt ttattgatta attgtgtcgc atatttatta
     1501 tcgttaaaaa tgcaattgac aaaagcgtaa attttaaggc tctaatgttt tttcttttgg
     1561 tatgttatag tcaagcaaag atgtattaag ttgcaaaagc atggaattga tttacgacat
     1621 ccgttcaact ttatgaccga gttgtcgctg tgtacttact ctttttttac tatgaaaagt
     1681 ttcaaacata atatcattaa ttatttcttt ttaaacgaag taacctgtaa cataaataat
     1741 aggggaaaag aagaaaaaaa ccataaaaat tcgagaatcg taacaattat tattgtaacg
     1801 atgacaaccc gattctttga gagagccaga gacatgactt ttgcattacc gtaattggtt
     1861 tctacttatg aattgtaaac ttgtgtaggg aatttcatcc accagctatc gcgggctgct
     1921 ctctgcatca gtgttagcac attttatatt aatgaagatt ttgctatgca aaatacattg
     1981 acaaggagga gggcaccaca caaaaagtta ggtgtaacag aaaatcatga aactatgatt
     2041 cctaatttat atattggagg attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca
     2101 ctcaatgacc tgaccatttg atggagttta agtcaatacc ttcttgaacc atttcccata
     2161 atggtgaaag ttccctcaag aattttactc tgtcagaaac ggccttaacg acgtagtcga
     2221 tcgacaagct tgcatgccct acaacaacta agaaaatggc tatcatgcgg aagctggtga
     2281 gaagaacagc atcgggacaa gggaaggaag aacaaagaca aagaaaacaa aagaaagcaa
     2341 ttgaaaacaa aacaaaacaa ttttcattcc ttctcttatc attccttttc ttttcttttc
     2401 tctcattcaa cgcactccat cgtatccgta ttcctcttat tttttctctt tctctatatc
     2461 catttctttc tctctaggtg tgtcctctct ctctcttcaa tttctctact ccgcattcca
     2521 acgcatcctt cccccaacct cccatttcct ccttacggcc cgatagcgat cgtctttccc
     2581 tcgctatcac tcgctaccgg cccctcctct gcaccgtaac ctcctacgta tttaccatat
     2641 cataaagttt tttccgacgc ttatcgctga ccccctgtcg ccctcctatt ggcttccgga
     2701 ttatcttctt gtccataagg tgatccatgc ttcctgaaga ttcccgaaat gtgtccactt
     2761 tggcggggaa tcattccatc cacttctttc tctctcgctt tcctcattcg gcgctcccct
     2821 tccgcgtctc attggtcttc cgctccgttt ttgctttgcc gatgttactt ggggagaggt
     2881 gcgataatcc tttcgcaaaa actcggtttg acgcctccca tggtataaat agtgggtggt
     2941 ggacaggtgc cttcgctttt cttttcttta agcaagagaa ttcctgcagg tcgactctag
     3001 aggatctcat atgacaacca gtcaccagcc tcaggacaga tacaaagctg tctggcttat
     3061 cttcttcatg ctgggtctgg gaacgctgct cccgtggaat tttttcatga cggccactca
     3121 gtatttcaca aaccgcctgg acatgtccca gaatgtgtcc ttggtcactg ctgaactgag
     3181 caaggacgcc caggcgtcag ccgcccctgc agcacccttg cctgagcgga actctctcag
     3241 tgccatcttc aacaatgtca tgaccctatg tgccatgctg cccctgctgt tattcaccta
     3301 cctcaactcc ttcctgcatc agaggatccc ccagtccgta cggatcctgg gcagcctggt
     3361 ggccatcctg ctggtgtttc tgatcactgc catcctggtg aaggtgcagc tggatgctct
     3421 gcccttcttt gtcatcacca tgatcaagat cgtgctcatt aattcatttg gtgccatcct
     3481 gcagggcagc ctgtttggtc tggctggcct tctgcctgcc agctacacgg cccccatcat
     3541 gagtggccag ggcctagcag gcttctttgc ctccgtggcc atgatctgcg ctattgccag
     3601 tggctcggaa ctatcagaaa gtgccttcgg ctactttatc acagcctgtg ctgttatcat
     3661 tttgaccatc atctgttacc tgggcctgcc ccgcctggaa ttctaccgct actaccagca
     3721 gctcaagctt gaaggacccg gggagcagga gaccaagttg gacctcatta gcaaaggaga
     3781 ggagccaaga gcaggcaaag aggaatctgg agtttcagtc tccaactctc agcccaccaa
     3841 tgaaagccac tctatcaaag ccatcctgaa aaatatctca gtcctggctt tctctgtctg
     3901 cttcatcttc actatcacca ttgggatgtt tccagccgtg actgttgagg tcaagtccag
     3961 catcgcaggc agcagcacct gggaacgtta cttcattcct gtgtcctgtt tcttgacttt
     4021 caatatcttt gactggttgg gccggagcct cacagctgta ttcatgtggc ctgggaagga
     4081 cagccgctgg ctgccaagcc tggtgctggc ccggctggtg tttgtgccac tgctgctgct
     4141 gtgcaacatt aagccccgcc gctacctgac tgtggtcttc gagcacgatg cctggttcat
     4201 cttcttcatg gctgcctttg ccttctccaa cggctacctc gccagcctct gcatgtgctt
     4261 cgggcccaag aaagtgaagc cagctgaggc agagaccgca ggagccatca tggccttctt
     4321 cctgtgtctg ggtctggcac tgggggctgt tttctccttc ctgttccggg caattgtgtg
     4381 agagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg
     4501 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt
     4561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctcggccccc
     4861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4921 aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     5041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5281 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5641 gtctatttcg ttcatccata gttgcctgac tgcccgtcgt gtagataact acgatacggg
     5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6001 ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6601 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
