LOCUS       sequence-433690-        5503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduction of a cyanamide-inducible I-SceI expression cassette, 
            use in combination with pITEdv1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5503)
  AUTHORS   McDonnell L, Evans S, Lu Z, Suchoronczak M, Leighton J, Ordeniza E, 
            Ritchie B, Valado N, Walsh N, Antoney J, Wang C , Scott C, Speight 
            R, Vickers CE, Peng B
  TITLE     Cyanamide-inducible expression of homing nuclease I-SceI for 
            iterative genome engineering and parallel promoter characterisation 
            in Saccharomyces cerevisiae
  JOURNAL   bioRxiv 2024.03.26.586389
REFERENCE   2  (bases 1 to 5503)
  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: "bioRxiv 
            2024.03.26.586389"
COMMENT     Sequence Label: pITESceIcmd
FEATURES             Location/Qualifiers
     source          1..5503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        249..592
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(808..824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(808..824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(821..843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    832..848
                     /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(856..862,863..880,881..886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    901..922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1039..1056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1210..1798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1290..1309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1969..2760,2761..2829))
                     /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     2592..2611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2830..2934)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3002..3020
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3058..3080)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3180..3199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3393..3415
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3407..3424
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3408..3424
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tctcagttaa aggaccagag tgtataaaat gtggcggaat cttacagttt gatgctgtcc
       61 gcgggcctca taagagttgt ggtaacaacg caggtgcgcg catctgctaa aatgtgtata
      121 ttagtttaaa aagttgtatg taataaaagt aaaatttaat attttggatg aaaaaaaagt
      181 tacgctaggg ataacagggt aatatagctg tttagcttgc ctcgtccccg ccgggtcacc
      241 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
      301 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
      361 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga
      421 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
      481 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
      541 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa
      601 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
      661 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
      721 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
      781 tgtttaaacg catgcggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2941 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3001 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     3061 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     3181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     3241 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     3301 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     3361 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     3421 cagtgcatgc gtttaaacca cgaaaagttc accataactt cgaataaagt cgcggaaaaa
     3481 agtaaacagc tattgctact caaatgaggt ttgcagaagc ttgttgaagc atgatgaagc
     3541 gttctaaacg cactattcat cattaaatat ttaaagctca taaaattgta ttcaattcct
     3601 attctaaatg gcttttattt ctattacaac tattagctct aaatccatat cctcataagc
     3661 agcaatcaat tctatctata ctttaagcgg ccgctttgaa agatgatact ctttatttct
     3721 agacagttat atatatatat atatatatat atatatatat atactcgagt tattacttaa
     3781 ggaaagtttc ggaggagata gtgttcggca gtttgtacat catctgcggg atcaggtacg
     3841 gtttgatcag gttgtagaag atcaggtaag acatagaatc gatgtagatg atcggtttgt
     3901 ttttgttgat ttttacgtaa cagttcagtt ggaatttgtt acgcagaccc ttaaccaggt
     3961 attctacttc ttcgaaagtg aaagactggg tgttcagtac gatcgatttg ttggtagagt
     4021 ttttgttgta atcccattta ccaccatcat ccatgaacca gtatgccaga gacatcgggg
     4081 tcaggtagtt ttcaaccagg ttgttcggga tggttttttt gttgttaacg atgaacaggt
     4141 tagccagttt gttgaaggct tggtgtttga aagtctgggc gccccaggtg attaccaggt
     4201 tacccaggtg gttaacacgt tcttttttgt gcggcgggga cagtacccac tgatcgtaca
     4261 gcagacatac gtggtccatg tatgctttgt ttttccactc gaactgcata cagtaggttt
     4321 taccttcatc acgagaacgg atgtaagcat cacccaggat cagaccgata cctgcttcga
     4381 actgttcgat gttcagttcg atcagctggg atttgtattc tttcagcagt ttagagttcg
     4441 gacccaggtt cattacctgg tttttttgat gcattttact agtgattgat tcttttgaag
     4501 agaagcaagg caaaaaacag tacgaagcaa cttcatgtta acagaagaag aaggaaacta
     4561 acccatttat atataatttt atgaggggac cgatcctaaa aagttatttg taaagctcgt
     4621 acaagattga ttgatgactt cccgacaaac tattgtgcgg tggagtattt tgtattatag
     4681 aaatgatagt gtccatgcac tttaataaga tcttttttgc ggacatttgt ttcggggccg
     4741 tccaaagttg tttgatggta taactccgca tatttgacat ctttctcttt taaaccagta
     4801 tcggttagtc ttttgcaatt ttcttgaggg attcagtcta gaaagttgat tttcattgct
     4861 gttaagataa aaattccttc taacccagct tgtactccgt atactttcaa tgctctcaaa
     4921 aagatatgga tactagttag tttagaccct gtagtttcac cagctgctag tattcaacgt
     4981 tcatgcgact tctataaagg agtgcttggc tttgatgtta caacattcgg gaactatcgc
     5041 agggctttgt ctttttttgg agtcagaaaa atcaatttgc atgaggtggg aaaggaattt
     5101 gagcccaagg cacagaagcc aactctcgga tctgtcgatc tgtgttttat cttagaggat
     5161 cctttgaata acagttaccc atcttgaaag cttggatttt ccaattgaag agggcactgt
     5221 aatccgtaca ggcgcagtca accctattat cagtatacat ccgtgattaa gatgaaagtg
     5281 tgatggaaat atcaaataga ataaaagaaa cggaatagag ctattttttt ttttatatta
     5341 tagacatagt ctaagcgagt ttaacctttg aaatataaaa aaaaacaaaa ggaaaaaagt
     5401 agtatgtggg ctgcactaaa ctaaaaccgt gaggtattta gatatatatt ctacattgtt
     5461 ttacgttata gcatgtataa agagctccag cttttgttcc ctt
//
