LOCUS       sequence-12706-e        5181 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 5181)
  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 5181)
  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: pXP320
FEATURES             Location/Qualifiers
     source          1..5181
                     /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    complement(440..473)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        598..785
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             786..1448
                     /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"
     protein_bind    complement(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"
     promoter        4293..4706
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      4744..4991
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4760..4778)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5154..5172
                     /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 tagcatacat tatacgaagt tatcgtttta
      481 agagcttggt gagcgctagg agtcactgcc aggtatcgtt tgaacacggc attagtcagg
      541 gaagtcataa cacagtcctt tcccgcaatt ttctttttct attactcttg gcctcctcta
      601 gtacactcta tattttttta tgcctcggta atgattttca tttttttttt tccacctagc
      661 ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat
      721 aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg
      781 caaagatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc aagattcaga
      841 ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc ccagaaaaag
      901 aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc cacacaggta
      961 tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg tcgctaatcg
     1021 ttgagtgcat tggtgactta cacatagacg accatcacac cactgaagac tgcgggattg
     1081 ctctcggtca agcttttaaa gaggccctag gggccgtgcg tggagtaaaa aggtttggat
     1141 caggatttgc gcctttggat gaggcacttt ccagagcggt ggtagatctt tcgaacaggc
     1201 cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc tcttgcgaga
     1261 tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc cacgttgatt
     1321 gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct cttgcggttg
     1381 ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc aaaggtgttc
     1441 ttatgtagtg acaccgatta tttaaagctg cagcatacga tatatataca tgtgtatata
     1501 tgtataccta tgaatgtcag taagtatgta tacgaacagt atgatactga agatgacaag
     1561 gtaatgcatc attctatacg tgtcattctg aacgaggcgc gctttccttt tttctttttg
     1621 ctttttcttt ttttttctct tgaactcgaa taacttcgta tagcatacat tatacgaagt
     1681 tatcccgggt accgagctcg aattcatcac gtgctataaa aataattata atttaaattt
     1741 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
     1801 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
     1861 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
     1921 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
     1981 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
     2041 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
     2101 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     2161 ttccatcatt aaaagatacg aggcgcgtgt aagttacgaa 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 tattaccgcg
     4261 aatccttaca tcacacccaa tcccccacaa gtgatccccc acacaccata gcttcaaaat
     4321 gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact
     4381 tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag ggtgtcgtta
     4441 attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt
     4501 cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt tttttttgat
     4561 ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc ttcaatttct
     4621 caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt gctcattaga
     4681 aagaaagcat agcaatctaa tctaagtttt aattacaaaa ctagtgatat ctgcgcactc
     4741 gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     4801 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     4861 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     4921 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     4981 ctttaatttg cggccaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5101 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5161 gtatcacgag gccctttcgt c
//
