LOCUS       sequence-210876-        8315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Same as pREP42, with better MCS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8315)
  TITLE     various
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8315)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pREP42-MCS+
FEATURES             Location/Qualifiers
     source          1..8315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        531..1685
                     /label=nmt1 P41 promoter
                     /note="mutant nmt1 promoter from Schizosaccharomyces pombe,
                     conferring medium strength thiamine-repressible expression"
     rep_origin      2802..3585
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(3935..3952)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3935..3951)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3944..3966)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4164..4619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4251..4270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4461..4482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4636..4655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4755..4777
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4815..4833)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4901..5005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5006..5074,5075..5866)
                     /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     complement(5224..5243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6037..6625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6526..6545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6779..6796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6913..6934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6949..6954,6955..6972,6973..6979)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6987..7003
                     /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)."
     primer_bind     6992..7014
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7011..7027
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7011..7027
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(7476..8270)
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
ORIGIN
        1 ttcttggtag gacaatacgg taagaaaaca cgacatgtgc agagatgccg acgaagcata
       61 gttaaactgg gatggtaaaa tcaattaaga atttataaag acaaaattgt ataagtctct
      121 aaaacatctt aattatacct cacagaacta tctaaaatat attcacaaag tgcaaacatt
      181 atcatgaaaa agaaccattt taatttaaag caagggcatt aaggcttatt tacagaattt
      241 cttacttttg taaagattat aaggctgatt atctttttca ccatgccaaa aattacacaa
      301 gatagaatgg atgtttgaaa ttaaacgtga gtatacaaac aaatacacta ggtaaatcga
      361 aacatttttt tctccattaa gtaacaaatt cctatttaga gaaagaatgc tgagtagatt
      421 aaataatcta tacaaacttt tttaacacaa atgcatacat atagccagtg ggatttgtag
      481 ctaagcttgc atgcctgcag gtcgatcgac tctagaggat cagaaaatta tcgccataaa
      541 agacagaata agtcatcagc ggttgtttca tttcctatat ttttttttta tttttttatt
      601 ttttaataag ggaaaattta acgtctaagg atacagaaga ttgttagcac attaaagtaa
      661 taaaggctta agtagtaagt gccttagcat gttattgtat ttcaaaggac ataatctaaa
      721 ataataacaa tatcatttct cacaagttat tcaattttct tttttttttc taataatatc
      781 aagaatgtat tatttgtttg acataagtca actaatttat ttaatatgct ggattaatct
      841 tgcagacatg taaattaaca agttttagtc aaataacgtt gaagtttcaa tgaactcaaa
      901 taatttctct ttttttttat ataaccataa tctgatttat attttccgca gggatcaact
      961 gaagttatga catttggatt ggatcactta taaccttggt cgccaaataa tacaaaaatc
     1021 agcgttataa aacaaagaag gtttttgtta agaaattaat cctctttctt gataagaaag
     1081 ttgaaccgaa attgcagata ctgatatatg aaaataatac ccacaatttt gggaatagcg
     1141 caagcctcaa tttaaacaat aggtgaggac acatgataat gacctcaatg attgttagaa
     1201 gaaaagagcc tcattacaaa atcgaaaaat gaatggttgg gtacaagttt ccaaaacatg
     1261 gtaaagtgga ctttgcgtat gagacgtaaa tagaaaaaaa cacttgttat atgttttcta
     1321 gaattattgt tgtctcttta tggttggatg atgcaaaata gtaatttcgg ttagttgctg
     1381 taaaacacca cgagacaaat agatatggat atttattaaa tcaggaaaaa cgtaactctc
     1441 ggctactgga tggttcagtc acccaacgat tactggggag agaaaacagg gcaaaagcaa
     1501 agcttaaagg aatccgattg tcattcggca atgtgcagcg aaactaaaaa ccggataatg
     1561 gacctgttaa tcgaaacatt gaagataaag gaagaggaat cctggcatat catcaattga
     1621 ataagttgaa ttaattattt caatctcatt ctcactttct gacttatagt cgctttgtta
     1681 aatcatatgc tcgagagatc tgtcgaccca tgggctagcg gatccccggg taaaaggaat
     1741 gtctcccttg ccagtactgc tagggttttt ctttcaaact atggaagccc attcaagctg
     1801 catattacga ttttgttttt cgcttttaga aagtggttta gatgagataa tagaaaaatt
     1861 cttgatctcc gacaacgagt acttttattt tttttgctaa tcactttact caatattagc
     1921 tcgaaatcgt agaaacgtag acgggtgcgg gataccgagt ggtgtagtta agaattttta
     1981 taaaccacgt ggcccaaaaa tatgaaccca aaacgtttat acatgagtat actttaagaa
     2041 ggctataccc cttcgtgtta gatgtagttt tagctaccca acccgagtct atgagcttga
     2101 cttcagatgt agaaggcatt aaatcgtttt gaatattaat taaaaaacga tgaaaattaa
     2161 atatttaaaa gcaatcatac gctgaaaatt tagtgctgtg gctaatcctt caacatggaa
     2221 atgccataaa agtgactttg acaaaaaaaa aagtatatac aggtagtaaa ctcatctact
     2281 tcattgactt tgtttacagc atgtggaagg aggaatattt attgctaaat cgtagtttaa
     2341 cattcaataa gtaatactat tgaaattcga caagattggc cgcatggatg aaaaagaggc
     2401 attttgcttt gggagaatta gtccaaatta gaactgaaaa aaaaaacttt acgaggcaaa
     2461 aatgtcggat tgagatcgta aaagttcgct cgtcgtcttt tgctttgtga ttgttttcat
     2521 ggatacatct tgctggatat ttaaatttta gtactatgta taagatattc tataaatgtt
     2581 ttatcaccca aacctgttag cgccttctta attctattca atctggcttt tgctctgaga
     2641 ctacttcttg gactttcact acttgttagt tatacggaat ttgtgtaatt agaagtgaaa
     2701 taatcctttc tattagtaat gcgagctcga attcgagtct aactccttaa ccactcggac
     2761 atagtgactt atctgacact cattgtaaat taatatatat ataggcattt tgtttagtta
     2821 aaggtactta agtaattagt ataaacgaac caattttata atcaggaagt taagtgaatg
     2881 gtagcacatg tcgtaaaaat tgtgaatttt tattgaataa tattttaaat acaagccttt
     2941 ctagactagg tatactcata aacatatatg agcaaaagga tagaggagat tacattgcat
     3001 cttctacaaa tttatttatt gccctttact gaaaaattaa ataatgagta ctaaatgata
     3061 aaaagcgctc agtacaagaa agattgcaaa aatattgcat tcttcatgaa ttaaagttgc
     3121 atataaggca tattgaaagt aatagtacta aaacagcagt tagcgaaaat taatagaatt
     3181 atattcgcaa gacaattgtg acattaaatt aaaaaattgt agaattttta ctatcctctt
     3241 taacgccatg agcctttata aaaaggttaa attagtttta acattctttt tttgagtaag
     3301 agtttacaat ttatcaaaac ctgtgttatt atattcatta gtttcaattt attagcatct
     3361 agagaaaaat caattggcag ttacattgtt ggaatttatg aagaaaaaga atctacaacg
     3421 gagaataagt tgctgatcgc tttccctaaa attgtatatt ttgctgagct tattttgaca
     3481 tttcgttgaa gttttctcta attcgcattc attttaagta aaacaatgag aaataaaatt
     3541 acaaaaaata caaattaaaa tacaattttt agctataatt atagacgatg cccttgtatc
     3601 ccattctgtc tcgcttgccc ctacttttta tcttttatat accataatga acgctgccgc
     3661 tactaaccat accccgattt tacatttcgg actcccaagg acgtacaaaa tagaaaacta
     3721 tagaaaaaaa taatcagaaa atagcatgtc atctctttgt aaaacgcgtt tgcaagaaga
     3781 aaggaaacaa tggagaagag atcatccatt tgtatgtaaa ttttagtaaa cttgaagaaa
     3841 tcactaacaa cttctcttac ttagggattc tatgcaaaac cttgtaaatc atctgatgga
     3901 ggactcgatt taatgaattg gaaggttgga attcactggc cgtcgtttta caacgtcgtg
     3961 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     4021 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     4081 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4141 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4201 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4261 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4321 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4381 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4441 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4501 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4561 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4621 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4681 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4741 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4801 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4861 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4921 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4981 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5101 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5161 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5221 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5281 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5401 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5581 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5881 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5941 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6001 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6061 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6661 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6721 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6781 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6841 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6901 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6961 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7021 ctatgaccat gattacgcca agcttgtgat attgacgaaa ctttttgaca tctaatttat
     7081 tctgttccaa caccaatgtt tataaccaag ttttatcttg tttgtctaca tggtatttta
     7141 cattcatcta catacatctt tcattggctt tgtacatagt tatcattaca agtctaaaaa
     7201 aattcactct tttcttattc aatgtcaatc caagagaaaa gattgtggta atgttgtagg
     7261 agcatgttta ataaattact atagcaaatt actttttatt cccaaggtgt ttatctataa
     7321 tagttaatat tttagtcgct acataaaatt ttaccaaaga gtacttgtat actaattcta
     7381 aatgccttct gacataaaac gcctaggaaa acaaacgcaa acaaggcatc gactttttca
     7441 ataaccaacc aaaaaaattt tacattagtc tttttttaat gctgagaaag tctttgctga
     7501 tatgccttcc aaccagcttc tctatatctc ttggcttcga caacaggatt acgaccagct
     7561 ccatagactc cacgaccaac aatgatgata tcgctaccgc agtttacaat cacttcttca
     7621 ggagtacgat attgctgtcc cagcccgtct cctttaacat ccaagccgat accaggggac
     7681 atagttatgt agtcgctttg aaggttagga aatcgacgac cagctataaa gccaaagcaa
     7741 aaatcggtat gcttctcaaa ccattctaag gttttctctg tgtaggaacc agtagccaaa
     7801 gagcctttgg aagacatttc agccaaaagc aagagaccac gtcccaaagg taaaccaact
     7861 tctttgaggc cttgtataat accctcgcct ggcactgtat ggcaatttgt gatatgagcc
     7921 caagaagcaa ttttgtacac accagatgca tattgtagct tgacggtatt tccaatgtct
     7981 gcgaatttgc gatcctcaaa gataagaaaa cgatgctttt tacctaaggc caccagtttt
     8041 tctaccatat cctggtcgaa atcctcgaca acgtcaatat gtgtcttgat aacacagaca
     8101 tagggtccaa ttttatctac caattctaag atttcggatt tcttcgtcaa atcgaccgcg
     8161 actgacaagt tgctttgctt ttcttccatc aaagccaaca attccttggc aatgggattt
     8221 ttcatcccct cagctctagc tgaatagctt tgaaatactc tagcatccat aactttgctt
     8281 ttaaaccttt aatttcgatc caagcaaaaa agagg
//
