LOCUS       sequence-12700-e        6286 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 6286)
  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 6286)
  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: pXP220
FEATURES             Location/Qualifiers
     source          1..6286
                     /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)."
     rep_origin      1710..3052
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2042..2089
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(3068..3084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3068..3084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3081..3103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3092..3108
                     /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(3116..3122,3123..3140,3141..3146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3161..3182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3299..3316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3470..4058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3550..3569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4229..5020,5021..5089))
                     /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     4852..4871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(5114..5361)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5327..5345
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6259..6277
                     /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 aattcgtatg atccaatatc aaaggaaatg atagcattga
     1741 aggatgagac taatccaatt gaggagtggc agcatataga acagctaaag ggtagtgctg
     1801 aaggaagcat acgatacccc gcatggaatg ggataatatc acaggaggta ctagactacc
     1861 tttcatccta cataaataga cgcatataag tacgcattta agcataaaca cgcactatgc
     1921 cgttcttctc atgtatatat atatacaggc aacacgcaga tataggtgcg acgtgaacag
     1981 tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc gctcgttttc ggaaacgctt
     2041 tgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt
     2101 gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc aaaaacgcac cggactgtaa
     2161 cgagctacta aaatattgcg aataccgctt ccacaaacat tgctcaaaag tatctctttg
     2221 ctatatatct ctgtgctata tccctatata acctacccat ccacctttcg ctccttgaac
     2281 ttgcatctaa actcgacctc tacatttttt atgtttatct ctagtattac tctttagaca
     2341 aaaaaattgt agtaagaact attcatagag tgaatcgaaa acaatacgaa aatgtaaaca
     2401 tttcctatac gtagtatata gagacaaaat agaagaaacc gttcataatt ttctgaccaa
     2461 tgaagaatca tcaacgctat cactttctgt tcacaaagta tgcgcaatcc acatcggtat
     2521 agaatataat cggggatgcc tttatcttga aaaaatgcac ccgcagcttc gctagtaatc
     2581 agtaaacgcg ggaagtggag tcaggctttt tttatggaag agaaaataga caccaaagta
     2641 gccttcttct aaccttaacg gacctacagt gcaaaaagtt atcaagagac tgcattatag
     2701 agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt tagaaaaata gcgctctcgg
     2761 gatgcatttt tgtagaacaa aaaagaagta tagattcttt gttggtaaaa tagcgctctc
     2821 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     2881 ctcgcgttgc atttttgttt tacaaaaatg aagcacagat tcttcgttgg taaaatagcg
     2941 ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta
     3001 gcgctctcgc gttgcatttt tgttctacaa aatgaagcac agatgcttcg ttgaattcgt
     3061 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3121 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3181 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3901 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5101 ttcaatattg gccgcaaatt aaagccttcg agcgtcccaa aaccttctca agcaaggttt
     5161 tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa atttgaaata
     5221 taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc tagacttcag
     5281 gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt gaatgtaagc
     5341 gtgacataac taattacatg actcgagtgc gcagatatca ctagtatatt tgttgtaaaa
     5401 agtagataat tacttccttg atgatctgta aaaaagagaa aaagaaagca tctaagaact
     5461 tgaaaaacta cgaattagaa aagaccaaat atgtatttct tgcattgacc aatttatgca
     5521 agtttatata tatgtaaatg taagtttcac gaggttctac taaactaaac cacccccttg
     5581 gttagaagaa aagagtgtgt gagaacaggc tgttgttgtc acacgattcg gacaattctg
     5641 tttgaaagag agagagtaac agtacgatcg aacgaacttt gctctggaga tcacagtggg
     5701 catcatagca tgtggtacta aaccctttcc cgccattcca gaaccttcga ttgcttgtta
     5761 caaaacctgt gagccgtcgc taggaccttg ttgtgtgacg aaattggaag ctgcaatcaa
     5821 taggaagaca ggaagtcgag cgtgtctggg ttttttcagt tttgttcttt ttgcaaacaa
     5881 atcacgagcg acggtaattt ctttctcgat aagaggccac gtgctttatg agggtaacat
     5941 caattcaaga aggagggaaa cacttccttt ttctggccct gataatagta tgagggtgaa
     6001 gccaaaataa aggattcgcg cccaaatcgg catctttaaa tgcaggtatg cgatagttcc
     6061 tcactctttc cttactcacg agtaattctt gcaaatgcct aatattattg aagcatttat
     6121 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6181 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6241 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
