LOCUS       sequence-428681-        2763 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 2763)
  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 2763)
  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: pPOM027_P4_tSynthGuo
FEATURES             Location/Qualifiers
     source          1..2763
                     /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(542..558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(542..558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(555..577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    566..582
                     /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(590..596,597..614,615..620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    635..656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(773..790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(944..1532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1024..1043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1703..2494,2495..2563))
                     /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     2326..2345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2564..2668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2736..2754
                     /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 tataactgtc tagaaataaa gagtatcatc tttcaaagct gtgagacctg agacgcatat
      481 gcacatcgga tgccgggacc gacgagtgca gaggcgtgca agcgagcttg gcgtaatcat
      541 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
      601 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
      661 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
      721 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
      781 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      841 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      901 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      961 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1021 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1081 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1141 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1201 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1261 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1321 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1381 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1441 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1501 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1561 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1621 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1681 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1741 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1801 gggagggctt accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg
     1861 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1921 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1981 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2041 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2101 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2161 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2221 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2281 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2341 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2401 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2461 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2521 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2581 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2641 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2701 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2761 gtc
//
