LOCUS       sequence-433683-        3690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Use together with pITEdv1, or pToxEXP1/pToxAmp plasmids to insert 
            heterologous gene(s) at ho locus. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3690)
  AUTHORS   Evans S, Lu Z, McDonnell L, Anderson W, Peralta F, Watkins T , Ahmed
            H, Luna-Flores CH, Loan T, Navone L, Trau M, Scott CH, Speight R, 
            Vickers CE, Peng B
  TITLE     The integration of tandem gene repeats via a bacterial type-II 
            toxin-antitoxin-mediated gene amplification (ToxAmp) system and 
            stability visualisation in Saccharomyces cerevisiae
  JOURNAL   bioRxiv 2024.01.30.578080
REFERENCE   2  (bases 1 to 3690)
  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 
            2024.01.30.578080"
COMMENT     Sequence Label: pJE13HD7
FEATURES             Location/Qualifiers
     source          1..3690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        532..875
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     primer_bind     complement(1091..1107)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1091..1107)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1104..1126)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1115..1131
                     /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(1139..1145,1146..1163,1164..1169))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1184..1205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1322..1339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1493..2081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1573..1592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2252..3043,3044..3112))
                     /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     2875..2894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3113..3217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3285..3303
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3341..3363)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3463..3482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(3676..3690,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3690,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgca tgcgtttaaa ccacgaaaag ttcaccataa cttcgaataa
       61 agtcgcggaa aaaagtaaac agctattgct actcaaatga ggtttgcaga agcttgttga
      121 agcatgatga agcgttctaa acgcactatt catcattaaa tatttaaagc tcataaaatt
      181 gtattcaatt cctattctaa atggctttta tttctattac aactattagc tctaaatcca
      241 tatcctcata agcagcaatc aattctatct atactttaaa agatctcagt taaaggacca
      301 gagtgtataa aatgtggcgg aatcttacag tttgatgctg tccgcgggcc tcataagagt
      361 tgtggtaaca acgcaggtgc gcgcatctgc taaaatgtgt atattagttt aaaaagttgt
      421 atgtaataaa agtaaaattt aatattttgg atgaaaaaaa agttacgcta gggataacag
      481 ggtaatatag ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
      541 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
      601 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
      661 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc ggacctgcga gcagggaaac
      721 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
      781 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
      841 tacagttctc acatcacatc cgaacataaa caaccatggg taaaaagcct gaactcaccg
      901 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
      961 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     1021 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttaa acgcatgcgg
     1081 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1141 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1201 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1261 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1321 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1381 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1441 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1501 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1561 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1621 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1681 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1741 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1801 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1861 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1921 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1981 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2041 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2101 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2161 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2221 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2281 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2341 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2401 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2461 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2521 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2581 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2641 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2701 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2761 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2821 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2881 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2941 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3001 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3061 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3121 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3181 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3241 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     3301 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3361 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3421 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3481 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3541 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3601 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3661 taacgccagg gttttcccag tcacgacgtt
//
