LOCUS       sequence-137516-        5987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  receiving vector of standard transcription units(TUs) in YeastFab 
            work, useful for further assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5987)
  AUTHORS   Guo Y, Dong J, Zhou T, Auxillos J, Li T, Zhang W, Wang L, Shen Y, 
            Luo Y, Zheng Y, Lin J, Chen GQ, Wu Q, Cai Y, Dai J
  TITLE     YeastFab: the design and construction of standard biological parts 
            for metabolic engineering in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2015 May 8. pii: gkv464.
  PUBMED    25956650
REFERENCE   2  (bases 1 to 5987)
  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: "Nucleic 
            Acids Res. 2015 May 8. pii: gkv464."
COMMENT     Sequence Label: POT9-RFP
FEATURES             Location/Qualifiers
     source          1..5987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..22)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    64..567
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        604..708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(709..777,778..1569)
                     /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(927..946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1740..2328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2229..2248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2482..2499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2652..2657,2658..2675,2676..2682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2690..2706
                     /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     2714..2730
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2714..2730
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2749..2769
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2751..2769
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2968..2989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3004..3009,3010..3027,3028..3034)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3042..3058
                     /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)."
     RBS             3076..3087
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3094..3771
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3816..3887
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3986..4005)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3987..4005)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4015..4032)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4015..4031)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4024..4046)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4176..4631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4263..4282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4473..4494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4762..5565)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5566..5786)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5812..5831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5931..5953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttagg
       61 acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
      121 gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
      181 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
      241 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
      301 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
      361 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
      421 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
      481 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
      541 attataatta tttttatagc acgtgatgaa aaggacccag gtggcacttt tcggggaaat
      601 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      661 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      721 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      781 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      841 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      901 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
      961 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1021 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1081 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1141 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1201 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1261 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1321 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1381 gctggctggt ttattgctga taaatctgga gccggtgagc gtggtagtcg cggtatcatt
     1441 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1501 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1561 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1621 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1681 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1741 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1801 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1861 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1921 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1981 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2041 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2101 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2161 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2221 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2281 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2341 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2401 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2461 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2521 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2581 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttacct cactcattag
     2641 gcaccccagg ctttacactt tatgcttccg gctcctatgt tgtgtggaat tgtgagcgga
     2701 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     2761 cactaaaggg aacaaaagct ggagctccac cgcggtggcg gccgcgaatt ccccgggtct
     2821 agaggtctcg gttggcagtg actcggtctc tgtcgggcta gagacggcaa tacgcaaacc
     2881 gcctctagcc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     2941 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     3001 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     3061 tcacacatac tagagaaaga ggagaaatac tagatggctt cctccgaaga cgttatcaaa
     3121 gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc
     3181 gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc
     3241 aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc
     3301 aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa
     3361 ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag
     3421 gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc
     3481 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
     3541 atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
     3601 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
     3661 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc
     3721 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataacgctga
     3781 tagtgctagt gtagatcgct actagagcca ggcatcaaat aaaacgaaag gctcagtcga
     3841 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagcgtct
     3901 cacctctgag agagacccaa gacactgcgg atcgagacca ctagtaccgg tctgcagctc
     3961 gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     4021 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4081 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4141 ccaacagttg cgcagcctga atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag
     4201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4561 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4621 gtttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4681 atagggtaat aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc
     4741 atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca
     4801 gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc
     4861 tcttccaaca ataataatgt cagatcctgt agacaccaca tcatccacgg ttctatactg
     4921 ttgacccaat gcgtcaccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc
     4981 gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt
     5041 gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt
     5101 gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc
     5161 ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc
     5221 ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc
     5281 agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg
     5341 cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca
     5401 aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa
     5461 tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg
     5521 actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt
     5581 cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc
     5641 aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt
     5701 ctgttcggag attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata
     5761 aaaaaaaaat gatgaattga attgaaaagc tgtggtatgg tgcactctca gtacaatctg
     5821 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5881 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacaatct ccgggagctg
     5941 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agattaa
//
