LOCUS       sequence-12694-e        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  AUTHORS   Fang F, Salmon K, Shen MW, Aeling KA, Ito E, Irwin B, Tran UP, 
            Hatfield GW, Da Silva NA, Sandmeyer S
  TITLE     A vector set for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2011 Feb;28(2):123-36. doi: 10.1002/yea.1824. Epub 2010 Oct 
            8.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 5431)
  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"; doi: "10.1002/yea.1824"; 
            journalName: "Yeast"; date: "2011-02"; volume: "28"; issue: "2"; 
            pages: "123-36"
COMMENT     Sequence Label: pXP120
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    440..473
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(675..1337)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(1338..1525)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     protein_bind    1650..1683
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2213..2229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2213..2229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2226..2248)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2237..2253
                     /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(2261..2267,2268..2285,2286..2291))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2306..2327
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2444..2461)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2615..3203)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2695..2714)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3374..4165,4166..4234))
                     /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     3997..4016
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(4259..4506)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4472..4490
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5404..5422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccggga taacttcgta taatgtatgc tatacgaagt tattcgagtt
      481 caagagaaaa aaaaagaaaa agcaaaaaga aaaaaggaaa gcgcgcctcg ttcagaatga
      541 cacgtataga atgatgcatt accttgtcat cttcagtatc atactgttcg tatacatact
      601 tactgacatt cataggtata catatataca catgtatata tatcgtatgc tgcagcttta
      661 aataatcggt gtcactacat aagaacacct ttggtggagg gaacatcgtt ggtaccattg
      721 ggcgaggtgg cttctcttat ggcaaccgca agagccttga acgcactctc actacggtga
      781 tgatcattct tgcctcgcag acaatcaacg tggagggtaa ttctgctagc ctctgcaaag
      841 ctttcaagaa aatgcgggat catctcgcaa gagagatctc ctactttctc cctttgcaaa
      901 ccaagttcga caactgcgta cggcctgttc gaaagatcta ccaccgctct ggaaagtgcc
      961 tcatccaaag gcgcaaatcc tgatccaaac ctttttactc cacgcacggc ccctagggcc
     1021 tctttaaaag cttgaccgag agcaatcccg cagtcttcag tggtgtgatg gtcgtctatg
     1081 tgtaagtcac caatgcactc aacgattagc gaccagccgg aatgcttggc cagagcatgt
     1141 atcatatggt ccagaaaccc tatacctgtg tggacgttaa tcacttgcga ttgtgtggcc
     1201 tgttctgcta ctgcttctgc ctctttttct gggaagatcg agtgctctat cgctagggga
     1261 ccacccttta aagagatcgc aatctgaatc ttggtttcat ttgtaatacg ctttactagg
     1321 gctttctgct ctgtcatctt tgccttcgtt tatcttgcct gctcattttt tagtatattc
     1381 ttcgaagaaa tcacattact ttatataatg tataattcat tatgtgataa tgccaatcgc
     1441 taagaaaaaa aaagagtcat ccgctaggtg gaaaaaaaaa aatgaaaatc attaccgagg
     1501 cataaaaaaa tatagagtgt actagaggag gccaagagta atagaaaaag aaaattgcgg
     1561 gaaaggactg tgttatgact tccctgacta atgccgtgtt caaacgatac ctggcagtga
     1621 ctcctagcgc tcaccaagct cttaaaacga taacttcgta taatgtatgc tatacgaagt
     1681 tatcccgggt accgagctcg aattcgtaac ttacacgcgc ctcgtatctt ttaatgatgg
     1741 aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga
     1801 aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta
     1861 aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta
     1921 aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg
     1981 gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa
     2041 gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa
     2101 aactattttt tctttaattt ctttttttac tttctatttt taatttatat atttatatta
     2161 aaaaatttaa attataatta tttttatagc acgtgatgaa ttcgtaatca tggtcatagc
     2221 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2281 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2341 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2401 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2461 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2521 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2581 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2641 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2701 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2761 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2821 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2881 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2941 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3001 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3061 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3121 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3181 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3241 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3301 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3361 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3421 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3481 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3541 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3601 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3661 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3721 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3781 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3841 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3901 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3961 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4021 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4081 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4141 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4201 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattggccgc
     4261 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
     4321 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
     4381 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
     4441 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
     4501 acatgactcg agtgcgcaga tatcactagt atatttgttg taaaaagtag ataattactt
     4561 ccttgatgat ctgtaaaaaa gagaaaaaga aagcatctaa gaacttgaaa aactacgaat
     4621 tagaaaagac caaatatgta tttcttgcat tgaccaattt atgcaagttt atatatatgt
     4681 aaatgtaagt ttcacgaggt tctactaaac taaaccaccc ccttggttag aagaaaagag
     4741 tgtgtgagaa caggctgttg ttgtcacacg attcggacaa ttctgtttga aagagagaga
     4801 gtaacagtac gatcgaacga actttgctct ggagatcaca gtgggcatca tagcatgtgg
     4861 tactaaaccc tttcccgcca ttccagaacc ttcgattgct tgttacaaaa cctgtgagcc
     4921 gtcgctagga ccttgttgtg tgacgaaatt ggaagctgca atcaatagga agacaggaag
     4981 tcgagcgtgt ctgggttttt tcagttttgt tctttttgca aacaaatcac gagcgacggt
     5041 aatttctttc tcgataagag gccacgtgct ttatgagggt aacatcaatt caagaaggag
     5101 ggaaacactt cctttttctg gccctgataa tagtatgagg gtgaagccaa aataaaggat
     5161 tcgcgcccaa atcggcatct ttaaatgcag gtatgcgata gttcctcact ctttccttac
     5221 tcacgagtaa ttcttgcaaa tgcctaatat tattgaagca tttatcaggg ttattgtctc
     5281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5341 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5401 aaaaataggc gtatcacgag gccctttcgt c
//
