LOCUS       sequence-415204-        5974 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 5974)
  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 5974)
  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: POT4-RFP
FEATURES             Location/Qualifiers
     source          1..5974
                     /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    63..566
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        603..707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(708..776,777..1568)
                     /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(926..945)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1739..2327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2228..2247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2481..2498
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2615..2636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2651..2656,2657..2674,2675..2681)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2689..2705
                     /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     2713..2729
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2713..2729
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2748..2768
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2750..2768
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2966..2987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3002..3007,3008..3025,3026..3032)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3040..3056
                     /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             3074..3085
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3092..3769
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3814..3885
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3984..4003)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3985..4003)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4013..4030)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4013..4029)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4022..4044)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4170..4625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4257..4276)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4467..4488
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4756..5559)
                     /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(5560..5775)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5799..5818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5918..5940
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
       61 cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg
      121 aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa
      181 gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa
      241 caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa atagatatac
      301 attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac
      361 agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta
      421 gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt
      481 ctttaatttc tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa
      541 ttataattat ttttatagca cgtgatgaaa aggacccagg tggcactttt cggggaaatg
      601 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      661 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      721 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      781 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      841 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      901 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      961 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1021 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1081 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1141 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1201 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1261 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1321 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1381 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggtagtcgc ggtatcattg
     1441 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1501 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1561 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1621 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1681 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1741 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1801 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1861 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1921 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1981 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2041 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2101 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2161 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2221 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2281 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2341 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2401 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2461 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2521 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2581 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2641 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2701 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     2761 actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgcgaattc cccgggtcta
     2821 gaggtctcgg tggcagtgac tcggtctcta ggcggctaga gacggcaata cgcaaaccgc
     2881 ctctagccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2941 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     3001 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     3061 acacatacta gagaaagagg agaaatacta gatggcttcc tccgaagacg ttatcaaaga
     3121 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
     3181 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
     3241 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
     3301 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
     3361 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
     3421 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
     3481 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
     3541 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
     3601 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
     3661 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
     3721 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat aacgctgata
     3781 gtgctagtgt agatcgctac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa
     3841 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagcgtctca
     3901 cctctgccag agacccaaga cactgcggat cgagaccact agtaccggtc tgcagctcga
     3961 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     4021 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4081 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4141 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     4201 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4261 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4321 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4381 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4441 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4501 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4561 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4621 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg
     4681 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
     4741 ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc
     4801 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
     4861 aacaataata atgtcagatc ctgtagacac cacatcatcc acggttctat actgttgacc
     4921 caatgcgtca cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
     4981 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
     5041 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
     5101 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
     5161 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
     5221 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
     5281 ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
     5341 tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
     5401 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     5461 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     5521 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     5581 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     5641 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     5701 ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa
     5761 aaatgatgaa ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca
     5821 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5881 ctcccggcat ccgcttacag acaagctgtg acaatctccg ggagctgcat gtgtcagagg
     5941 ttttcaccgt catcaccgaa acgcgcgaga ttaa
//
