LOCUS       sequence-428683-        2793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic terminator for gene expression in yeasts and fungi. Part 
            type 4 following the YeastToolkit MoClo grammar..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2793)
  AUTHORS   Hebra T, Smrckova H, Elkatmis B, Prevorovsky M, Pluskal T
  TITLE     POMBOX: A Fission Yeast Cloning Toolkit for Molecular and Synthetic 
            Biology.
  JOURNAL   ACS Synth Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529.
  PUBMED    37991801
REFERENCE   2  (bases 1 to 2793)
  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: "ACS Synth 
            Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529."
COMMENT     Sequence Label: pPOM028_P4_tSynth3
FEATURES             Location/Qualifiers
     source          1..2793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     313..335
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     327..344
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     328..344
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(572..588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(572..588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(585..607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    596..612
                     /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(620..626,627..644,645..650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    665..686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(803..820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(974..1562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1054..1073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1733..2524,2525..2593))
                     /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     2356..2375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2594..2698)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2766..2784
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
       61 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      121 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      181 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      241 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      301 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg gagatcggta
      361 cttcgcgaat gcgtcgagat ctagacatcc gtctcatcgg tctcaatcct aactcgagta
      421 tatatatata tatatatata tatatatata tataactgtc tagaaataaa gagtatcatc
      481 tttcaaagct gtgagacctg agacgcatat gcacatcgga tgccgggacc gacgagtgca
      541 gaggcgtgca agcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      601 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      661 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      721 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      781 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      841 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      901 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      961 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1021 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1081 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1141 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1201 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1261 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1321 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1381 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1441 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1501 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1561 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1621 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1681 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1741 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1801 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1861 caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1921 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1981 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2041 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2101 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2161 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2221 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2281 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2341 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2401 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2461 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2521 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2581 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2641 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2701 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     2761 ataaaaatag gcgtatcacg aggccctttc gtc
//
