LOCUS       sequence-332961-        6494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in S. pombe from adh81 promoter, integrated at arg3-D4 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6494)
  TITLE     Carr lab yeast expression plasmids
REFERENCE   2  (bases 1 to 6494)
  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"
COMMENT     Sequence Label: Padh81-MCS-TADHdh1-Sp.arg3+/pUC19*
FEATURES             Location/Qualifiers
     source          1..6494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(547..1338,1339..1407))
                     /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     1170..1189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1408..1512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1580..1598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1636..1658)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1758..1777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1969..1991
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1984..2000
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2013..2029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4331..5066
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     terminator      5184..5371
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5239..5263)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(5880..5896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5880..5896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5893..5915)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5904..5920
                     /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(5928..5934,5935..5952,5953..5958))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5973..5994
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6111..6128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6282..6494,1..376))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6362..6381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
       61 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
      661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
      721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
      781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
      841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
      901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
      961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1561 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     1621 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1681 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1741 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atgcggtgtg
     1801 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     1861 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     1921 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     1981 gttgtaaaac gacggccagt gaattcgcgg ttgtaaaacg acggccagtg aattcgcggc
     2041 cgcagcttgt aatatagcga atctgtgaac tcattgttca tttcatggta gttgttgatt
     2101 tcacaagcgt gctgagttat caatatagta acacaattgg ttttatttca gctcgcttga
     2161 atatagtgtc aagagataat tgaataaaca aaaaaaataa ctacacgttt tttagataca
     2221 ttttatttac aatgtattgg ttctttaaag aatgggtggt aaataaaaaa aatattacaa
     2281 ggtcgaaaaa aattcatccc ttttcattag ttctataatt aaatgctaaa cccatgaata
     2341 ttcttttact gctaatcaat taacgatttt ggtatttaaa agctattgct gataagtatt
     2401 ctgacgttct gtatcaatat taccatgaat aacataagaa aaagaaagga aatagaatgt
     2461 ttatgagtag tttttaattt ttgacttgtt ttatcatatt tcatcaacgt actagcttgt
     2521 ttgcaactga actctagtgt acgataatgt aaaaattttc catctcgtcg aaaatccgca
     2581 tacgatatat tacggcgcca atctcgcagt ctgagagaga actagatatc ggctatatgc
     2641 aatctcaccc tgaatatcta agaaattacc agtaacttgt gggttccttg tagtcgtcta
     2701 caaaaaccgt atttttgata tatttagtcg agaagaatta ttcagtttgc aattgaaaat
     2761 gtctttcaaa aaatttcctc gtcatttact atccattcgt gatcttagtc gcggagagat
     2821 tgtaaagttg atcgatcggt cttctgaaat taaacaagcc tataagcaga attttcagaa
     2881 tcgccgttct gttcaaatga gtggtttgtc gtctcagaat gtcgccatga tcttcagtaa
     2941 gcgtagcact cgtactcgtg tatcagtcga aagtgctgtc agttgccttg gcggcaatgc
     3001 catgttttta ggtaaagatg atatccaatt aggagtgaat gagtctttat acgatacttc
     3061 aaaagttata tcaagtatgg tttctggtat tgttgctcgc gtaaacaaat attcagatgt
     3121 tgctacgctc gctaaacatg cctcttgtcc tgttattaat ggtctttgtg acacgtttca
     3181 tcctctccag gcgttagcag atttactcac tattaaggaa actttcaaga gctttgacgg
     3241 tttaaaagtt gcatgggtag gtgatgccaa taacgtttta catgatttga tgattgcaaa
     3301 tgccaaggtt ggtattcata ctagcgttgc taagcccaag gacgtcaacg ttcgcgacga
     3361 tattttatca atcgttaacg aagctgcgaa cgaaaacggt tctacttttg aaattgttaa
     3421 cgaccccaag gttgctgtta aaaatgctga tatcgtcgtt accgacactt ggatttctat
     3481 gggtcaagaa gctgagaagg aacaacgtct taaacaattt acaggattcc aagtcaccgg
     3541 agagatcatg aaacttgcca aacctagctg caaattcatg cattgtttac cccgtcatcc
     3601 cgaggaagtg tctgacgagg tattttatgg agagaacagt ttagttttcc aagaagcaga
     3661 gaaccgcaaa tggaccactg ttgctgtctt ggaggctttg cttgtcaacc gtggtgaaat
     3721 tcttcctcct gcctcagcct aaactgaaat ccttaatttc atttttatct ggggaatgtg
     3781 caaacaatgc caactttata atttttttta ttgggcgtgc aaattgtgat attcattttt
     3841 gaaattttat gtactttgtt tcacaaaaag aaaattgact tttgagaaaa ttttttaatt
     3901 gacaattact tttcagattt tttatgcgca ggacacaggc ttctaaagac gtacttatac
     3961 gttaaaaaag gggatggatc aatttggctc attagtccaa caaggtttgt gtaaaattat
     4021 tacccgctta atgctatcct cttaactaga tatttcaagg tatcaagcaa tcatagtatt
     4081 aagcactatt atatgcaaaa tacaatttaa aattaataga aactacaaat aagcgcaaca
     4141 ctcataagga aaaaggtttg ggacaataaa ggtttcgtca atgcggaatc tacgacagaa
     4201 agaaaactta gtgtaggcat cgaaatttat tatatttcaa gcatgtatgc tagtaaaata
     4261 aaacgtaaac tcattgattc aagaaatcta tagaaaaaaa gctagcgtga cgcagacatt
     4321 cgaatggcat gccctacaac aactaagaaa atggctatca tgcggaagct ggtgagaaga
     4381 acagcatcgg gacaagggaa ggaagaacaa agacaaagaa aacaaaagaa agcaattgaa
     4441 aacaaaacaa aacaattttc attccttctc ttatcattcc ttttcttttc ttttctctca
     4501 ttcaacgcac tccatcgtat ccgtattcct cttatttttt ctctttctct atatccattt
     4561 ctttctctct aggtgtgtcc tctctctctc ttcaatttct ctactccgca ttccaacgca
     4621 tccttccccc aacctcccat ttcctcctta cggcccgata gcgatcgtct ttccctcgct
     4681 atcactcgct accggcccct cctctgcacc gtaacctcct acgtatttac catatcataa
     4741 agttttttcc gacgcttatc gctgaccccc tgtcgccctc ctattggctt ccggattatc
     4801 ttcttgtcca taaggtgatc catgcttcct gaagattccc gaaatgtgtc cactttggcg
     4861 gggaatcatt ccatccactt ctttctctct cgctttcctc attcggcgct ccccttccgc
     4921 gtctcattgg tcttccgctc cgtttttgct ttgccgatgt tacttgggga gaggtgcgat
     4981 aatcctttcg caaaaactcg gtttgacgcc tcccatggat gtgggtggtg gacaggtgcc
     5041 ttcgcttttc tttaagcaag agaattccat tgtcttgact atcacaaact tttaagtctt
     5101 ttcttttttc taaccacata catatgctgg catgcactcc cgggagagag ctccgtctgc
     5161 agcgagtcga cacttctaaa taagcgaatt tcttatgatt tatgattttt attattaaat
     5221 aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgaa
     5281 aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag gtatagtatg
     5341 aggtcgctct tattgaccac acctctaccg gcagatccgc tagggataac agggtaatat
     5401 actagtgatc tgggcaagta gagtgtgcat catcgaaatc cggcttcttt cataaactta
     5461 tcgtattgct ctttaagtgc tttgctttta tcaataaatg gctttttctt gtcttccgcc
     5521 aattccttcc acgcctttga tgcattttgg gtctcttcaa cgagtgtgga aatggcttct
     5581 ggaataccta actcttgacg tagcttggga ttgtttcgaa gctccttgta aaagaggata
     5641 aaagcagaag aaggtttttt aggaacatct ggatgacgaa gtctgtaata gcgactgccc
     5701 gtgctgtgag cctccttcga gcgacgacga ttttcactat cgatttctgt aggtgtaaag
     5761 gatcgtagag tctccataaa acgctttttc tcagcatcga actgctcctg gagtttgttt
     5821 ttatactttt ggatctcatc ggcagtcaaa gctgcggccg caagcttggc gtaatcatgg
     5881 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5941 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6001 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6061 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6181 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6481 tcacgctgta ggta
//
