LOCUS       sequence-138758-        7654 bp    DNA     circular SYN 15-APR-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 7654)
  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 7654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: POT5-TEF2p-ycrtYB-ADH1t
FEATURES             Location/Qualifiers
     source          1..7654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..103)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(85..103)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(113..130)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(113..129)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(122..144)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      274..729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(361..380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     571..592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(860..1663)
                     /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(1664..1884)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(1910..1929)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2029..2051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2089..2107)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2149..2652
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2689..2793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2794..2862,2863..3654)
                     /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(3012..3031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3825..4413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4314..4333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4567..4584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4737..4742,4743..4760,4761..4767)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4775..4791
                     /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     4780..4802
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4799..4815
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4799..4815
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4834..4854
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4836..4854
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      7468..7654
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(7522..7546)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
ORIGIN
        1 cctctgagag agacccagga cactgcggat cgagaccact agtaccggtc tgcagctcga
       61 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
      121 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
      181 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      241 aacagttgcg cagcctgaat ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg
      301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      481 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      601 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
      721 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      781 agggtaataa ctgatataat taaattgaag ctctaatttg tgagtttagt atacatgcat
      841 ttacttataa tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc
      901 ctgcttttct gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc
      961 ttccaacaat aataatgtca gatcctgtag acaccacatc atccacggtt ctatactgtt
     1021 gacccaatgc gtcacccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt
     1081 aaccttcatc tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt
     1141 cgctcttcgc aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc
     1201 atgacaattc tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct
     1261 gcttcaaacc gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc
     1321 attctgctat tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag
     1381 caaattttct gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct
     1441 taactgtgcc ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa
     1501 ttttgggacc taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg
     1561 aagcacacaa gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac
     1621 taggatgagt agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc
     1681 tgcaggtttt tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa
     1741 cactacatat gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct
     1801 gttcggagat taccgaatca aaaaaatttc aaagaaaccg aaatcaaaaa aaagaataaa
     1861 aaaaaaatga tgaattgaat tgaaaagctg tggtatggtg cactctcagt acaatctgct
     1921 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     1981 gggcttgtct gctcccggca tccgcttaca gacaagctgt gacaatctcc gggagctgca
     2041 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag attaaagggc ctcgtgatac
     2101 gcctattttt ataggttaat gtcatgataa taatggtttc ttaggacgga tcgcttgcct
     2161 gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt
     2221 ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt
     2281 tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt
     2341 tacatatata tttattagac aagaaaagca gattaaatag atatacattc gattaacgat
     2401 aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac
     2461 aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga
     2521 tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt
     2581 tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt
     2641 atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     2701 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2761 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2821 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2881 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2941 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     3001 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     3061 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     3121 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3181 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3241 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3301 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3361 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3421 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3481 gctgataaat ctggagccgg tgagcgtggt agtcgcggta tcattgcagc actggggcca
     3541 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3601 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3661 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3721 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3781 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3841 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3901 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3961 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4021 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4081 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4141 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4201 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4261 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4321 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4381 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4441 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4501 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4561 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     4621 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     4681 ggcagtgagc gcaacgcaat taatgtgagt tacctcactc attaggcacc ccaggcttta
     4741 cactttatgc ttccggctcc tatgttgtgt ggaattgtga gcggataaca atttcacaca
     4801 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     4861 aagctggagc tccaccgcgg tggcggccgc gaattccccg ggtctagagg tctcggttgg
     4921 cagtgactcg gtctcttgcc ggctgggcgc cataaccaag gtatctatag accgccaatc
     4981 agcaaactac ctccgtacat tcatgttgca cccacacatt tatacaccca gaccgcgaca
     5041 aattacccat aaggttgttt gtgacggcgt cgtacaagag aacgtgggaa ctttttaggc
     5101 tcaccaaaaa aggaagaaaa aatacgagtt gctgacagaa gcctcaagaa aaaaaaaatt
     5161 cttcttcgac tatgctggag gcagagatga tcgagccggt agttaactat atatagctaa
     5221 attggttcca tcaccttctt ttctggtgtc gctccttcta gtgctatttc tggcttttcc
     5281 tatttttttt tttccatttt tctttctctc tttctaatat ataaattctc ttgcattttc
     5341 tatttttctc tctatctatt ctacttgttt attcccttca aggttttttt ttaaggagta
     5401 cttgttttta gaatatgcgg tcaacgaact ataattaact aaagatgacc gctttggctt
     5461 actaccaaat ccacttgatc tacaccttgc caatcttggg tttgttgggt ttgttgacct
     5521 ctccaatctt gaccaagttc gacatctaca aaatctctat cttggttttc atcgctttct
     5581 ctgctaccac cccatgggac tcttggatca tcagaaacgg tgcttggacc tacccatctg
     5641 ctgaatctgg tcaaggtgtt ttcggaacct tcttggacgt tccatacgaa gaatacgctt
     5701 tcttcgttat ccaaaccgtt atcacaggtt tggtttacgt tttggctacc agacacttgt
     5761 tgccatcttt ggctttgcca aagacccgat cttctgcttt gtctttggct ttgaaggctt
     5821 tgatcccatt gccaatcatc tacttgttca ccgctcaccc atctccatct ccagacccat
     5881 tggttaccga ccactacttc tacatgagag ctttgtcttt gttgatcacc ccaccaacca
     5941 tgttgttggc tgctttgtct ggtgaatacg ctttcgactg gaagtctggt agagctaagt
     6001 ctaccatcgc tgctatcatg atcccaaccg tttacttgat ctgggttgac tacgttgctg
     6061 ttggtcaaga ctcttggtct atcaacgacg aaaagatcgt tggttggaga ttgggtggtg
     6121 ttttgccaat cgaagaagct atgttcttct tgttgaccaa cttgatgatc gttttgggtt
     6181 tgtctgcttg tgaccacacc caggctttgt acttgttgca cggtagaacc atctacggta
     6241 acaagaagat gccatcttct ttcccattga tcaccccacc agttttgtct ttgttcttct
     6301 ctagtagacc atactcttct caaccaaaga gagacttgga attggctgtt aagttgttgg
     6361 aagaaaagtc tcgatctttc ttcgttgctt ctgctggttt cccatctgaa gttagagaaa
     6421 gattggttgg tttgtacgct ttctgtagag ttaccgacga cttgatcgac tctccagaag
     6481 tttcttctaa cccacacgct accatcgaca tggtttctga cttcttgacc ttgttgttcg
     6541 gtccaccatt gcacccatct caaccagaca aaatcttgtc ttctccattg ttgccaccat
     6601 ctcacccaag tagaccaaca ggtatgtacc cattgccacc accaccatct ttgtctccag
     6661 ctgaattggt tcaattcttg accgaaagag ttccagttca ataccacttc gctttcagat
     6721 tgttggctaa gttgcaaggt ttgatcccaa gatacccatt ggacgaattg ttgagaggtt
     6781 acaccaccga cttgatcttc ccattgtcta ccgaagctgt tcaagctaga aagaccccaa
     6841 tcgaaaccac cgctgacttg ttggactacg gtttgtgtgt tgctggttct gttgctgaat
     6901 tgttggttta cgtttcttgg gcttctgctc catctcaagt tccagctacc atcgaagaaa
     6961 gagaagctgt tttggttgct agtagagaaa tgggaaccgc tttgcaattg gttaacatcg
     7021 ctagagacat caagggtgac gctaccgaag gtagattcta cttgccattg tctttcttcg
     7081 gtttgcgcga cgaatctaag ttggctatcc caaccgactg gaccgaacca agaccacaag
     7141 acttcgacaa gttgttgtct ttgtctccat cttctacctt gccatcttct aacgcttctg
     7201 aatctttcag attcgaatgg aagacctact ctttgccatt ggttgcttac gctgaagact
     7261 tggctaagca ctcttacaag ggtatcgaca gattgccaac cgaagttcaa gctggtatga
     7321 gagctgcttg tgcttcttac ttgttgatcg gtagagaaat caaggttgtt tggaagggtg
     7381 acgttggtga aagaagaacc gttgctggtt ggagaagagt tagaaaggtt ttgtctgttg
     7441 ttatgtctgg ttgggaaggt caatagccga atttcttatg atttatgatt tttattatta
     7501 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     7561 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     7621 atgaggtcgc tcttattgac cacacctcta ccgg
//
