LOCUS       sequence-110587-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TU for combinatorial construction of ?-carotene pathway.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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: POT2-CYC1p-ycrtE-ADH1t
FEATURES             Location/Qualifiers
     source          1..6559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(91..111)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(91..109)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(130..146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(130..146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(143..165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    154..170
                     /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(178..184,185..202,203..208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(361..378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(532..1120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(612..631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1291..2082,2083..2151))
                     /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     1914..1933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2152..2256)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2293..2796
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2838..2856
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2894..2916)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3016..3035
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3061..3281
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3282..4085
                     /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"
     rep_origin      complement(4216..4671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4353..4374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4565..4584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4801..4823
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4815..4832
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4816..4832
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4842..4861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4842..4860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4945..5131)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     5053..5077
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 agccaggtag agaccgagtc actgccaacc gagacctcta gacccgggga attcgcggcc
       61 gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
      121 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
      181 cataggagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga ggtaactcac
      241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
      301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      961 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gactaccacg
     1441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     2281 tgggtccttt tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata
     2341 tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg
     2401 aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct
     2461 gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa
     2521 atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt
     2581 tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt
     2641 ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
     2701 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
     2761 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc ctaagaaacc attattatca
     2821 tgacattaac ctataaaaat aggcgtatca cgaggccctt taatctcgcg cgtttcggtg
     2881 atgacggtga aaacctctga cacatgcagc tcccggagat tgtcacagct tgtctgtaag
     2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     3001 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat accacagctt
     3061 ttcaattcaa ttcatcattt tttttttatt cttttttttg atttcggttt ctttgaaatt
     3121 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     3181 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     3241 aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa gctacatata
     3301 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     3361 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     3421 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3481 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3541 tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3601 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3661 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3721 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3781 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3841 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3901 atgacaaggg tgacgcattg ggtcaacagt atagaaccgt ggatgatgtg gtgtctacag
     3961 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     4021 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     4081 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     4141 tttaattata tcagttatta ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4201 aaataccgca tcaggaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     4261 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4321 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4381 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4441 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4501 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4561 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4621 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcgcgccatt
     4681 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4741 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4801 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     4861 gcgaattggg taccgggccc cccctcgagc tgcagaccgg tactagtggt ctcgatccgc
     4921 agtgtcttgg gtctctgcct gaggccggta gaggtgtggt caataagagc gacctcatgc
     4981 tatacctgag aaagcaacct gacctacagg aaagagttac tcaagaataa gaattttcgt
     5041 tttaaaacct aagagtcact ttaaaatttg tatacactta ttttttttat aacttattta
     5101 ataataaaaa tcataaatca taagaaattc ggctacaatg ggatgtcagc caacttcttc
     5161 aagatagctt ccaacttaac gttcttaccg gttctacctt cagagtcaac gtcacccaag
     5221 tccattctac tgttagcttc agcgaactca ccttgcaatc tacccaattc tctttccaaa
     5281 gcaccagaca aggtgttcaa aacttcttgg gtgtattcga aagagtgggt ttcggttctc
     5341 atgtagttaa cacagtggtg caagatttct ggagaggtag acttcttttg caaggtgttg
     5401 ataaccaatc tactagatgg gttagcgtgg atagagtgga tggttgggaa agagaactta
     5461 ccttcggtca agtcttcagc gaagttcttg ttgtgagcgt attcagaaga ttgcaagttc
     5521 atgtagtcgt cgcggatttg gaagtagata gagatcaagt taaccaattg aacgaagtcg
     5581 atgtcacatt cagacttagc catcatcaat ctaacagcga ttctgaacaa accaccggtc
     5641 ttacccaaaa ccatcttaac gtattcttct tcagatggac aggtcaaaga gtctctccag
     5701 aacaattcca aaccttgacc tctgtgcaaa gacaagattt cgtcggtgat aaccttgtcc
     5761 aagtaggtgt ccttagagaa tgggatttga gagtttgggg tagaggttcc accgttttca
     5821 gaagaagcag aagaagaaga agaagcagaa gaaacagaag attgcaaaga agcagaagat
     5881 ggtgggataa ctggcattgg gattggggtt ggtctgagct tgaagatttc ttggtaagcc
     5941 aagaagtaaa cgtagttagc ggtgttgatg gtttgtggga taccgtagat caagtgagca
     6001 actggagaac ctcttctcaa aacagaagag tcttcaacgt cgtccatcaa caaagaaccg
     6061 gtgtgcaaca taccaacaac gttttggata acttccaagt cttccttctt aacgtccaac
     6121 cagtagttga aagcctcgat caattgagat ctgatttcct tacctgggtt cttacccaag
     6181 tagtggtatg gttccaacaa aacgatgtcg tcttgtgggg tgaactccaa tgggatagcg
     6241 gtcaagatgt tagcgtagtc catcttagtg tgtgtatttg tgtttgcgtg tctatagaag
     6301 tatagtaatt tatgctgcaa aggtcctaat gtataaggaa agaatattta gagaaaagaa
     6361 gaaaacaaga gttttatata catacagagc acatgcatgc catatgatca tgtgtcgtcg
     6421 cacacatata tatatgcctg tatgtgtcag cactaaagtt gcctggccat ccacgctata
     6481 tatacacgcc tggcggatct gctcgaggat tgcctacgcg tgggcttgat ccaccaacca
     6541 acgctcgcca aatgagctc
//
