LOCUS       sequence-428692-        2792 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 2792)
  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 2792)
  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: pPOM032_P4_tSynth30
FEATURES             Location/Qualifiers
     source          1..2792
                     /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(571..587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(571..587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(584..606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    595..611
                     /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(619..625,626..643,644..649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    664..685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(802..819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(973..1561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1053..1072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1732..2523,2524..2592))
                     /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     2355..2374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2593..2697)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2765..2783
                     /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 aactcgagtt
      421 tttttttata tatatatata tatatatata aactcattta cttatgtagg aataaatttt
      481 ttcaaagctg tgagacctga gacgcatatg cacatcggat gccgggaccg acgagtgcag
      541 aggcgtgcaa gcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
      721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
      781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1141 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1381 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1861 aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc
     1921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     2761 taaaaatagg cgtatcacga ggccctttcg tc
//
