LOCUS       sequence-138576-        5986 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 5986)
  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 5986)
  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: POT7-RFP
FEATURES             Location/Qualifiers
     source          1..5986
                     /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(3985..4004)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3986..4004)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4014..4031)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4014..4030)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4023..4045)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4175..4630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4262..4281)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4472..4493
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4761..5564)
                     /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(5565..5785)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5811..5830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5930..5952
                     /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 tcactggcta 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 tagacgtctc
     3901 acctctgaga gagacccaag acactgcgga tcgagaccac tagtaccggt ctgcagctcg
     3961 agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4021 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4081 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4141 caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc
     4201 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4261 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4321 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4381 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4441 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4501 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4561 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4621 tttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4681 tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     4741 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     4801 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     4861 cttccaacaa taataatgtc agatcctgta gacaccacat catccacggt tctatactgt
     4921 tgacccaatg cgtcaccctt gtcatctaaa cccacaccgg gtgtcataat caaccaatcg
     4981 taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac aaaatctttg
     5041 tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag ggagcccttg
     5101 catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc ttctgccgcc
     5161 tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt aatgtctgcc
     5221 cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt accaatgtca
     5281 gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc
     5341 ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt tagtaaacaa
     5401 attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg aacatccaat
     5461 gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc agcaacagga
     5521 ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta tcttcgtttc
     5581 ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca tgtttcttca
     5641 acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     5701 tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa
     5761 aaaaaaaatg atgaattgaa ttgaaaagct gtggtatggt gcactctcag tacaatctgc
     5821 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5881 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgacaatctc cgggagctgc
     5941 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gattaa
//
