LOCUS       sequence-12695-e        5696 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 5696)
  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 5696)
  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: pXP122
FEATURES             Location/Qualifiers
     source          1..5696
                     /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)."
     protein_bind    1915..1948
                     /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(2478..2494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2478..2494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2491..2513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2502..2518
                     /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(2526..2532,2533..2550,2551..2556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2571..2592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2709..2726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2880..3468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2960..2979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3639..4430,4431..4499))
                     /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     4262..4281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      complement(4524..4771)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4737..4755
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5669..5687
                     /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 tatggatttt
      481 cttaacttct tcggcgacag catcaccgac ttcggtggta ctgttggaac cacctaaatc
      541 accagttctg atacctgcat ccaaaacctt tttaactgca tcttcaatgg ccttaccttc
      601 ttcaggcaag ttcaatgaca atttcaacat cattgcagca gacaagatag tggcgatagg
      661 gttgacctta ttctttggca aatctggagc agaaccgtgg catggttcgt acaaaccaaa
      721 tgcggtgttc ttgtctggca aagaggccaa ggacgcagat ggcaacaaac ccaaggaacc
      781 tgggataacg gaggcttcat cggagatgat atcaccaaac atgttgctgg tgattataat
      841 accatttagg tgggttgggt tcttaactag gatcatggcg gcagaatcaa tcaattgatg
      901 ttgaaccttc aatgtaggaa attcgttctt gatggtttcc tccacagttt ttctccataa
      961 tcttgaagag gccaaaacat tagctttatc caaggaccaa ataggcaatg gtggctcatg
     1021 ttgtagggcc atgaaagcgg ccattcttgt gattctttgc acttctggaa cggtgtattg
     1081 ttcactatcc caagcgacac catcaccatc atcttccttt ctcttaccaa agtaaatacc
     1141 tcccactaat tctctgacaa caacgaagtc agtaccttta gcaaattgtg gcttgattgg
     1201 agataagtct aaaagagagt cggatgcaaa gttacatggt cttaagttgg cgtacaattg
     1261 aagttcttta cggattttta gtaaaccttg ttcaggtcta acactacctg taccccattt
     1321 aggaccaccc acagcaccta acaaaacggc atcagccttc ttggaggctt ccagcgcctc
     1381 atctggaagt gggacacctg tagcttcgat agcagcacca ccaattaaat gattttcgaa
     1441 atcgaacttg acattggaac gaacatcaga aatagcttta agaaccttaa tggcttcggc
     1501 tgtgatttct tgaccaacgt ggtcacctgg caaaacgacg atcttcttag gggcagacat
     1561 tagaatggta tatccttgaa atatatatat atattgctga aatgtaaaag gtaagaaaag
     1621 ttagaaagta agacgattgc taaccaccta ttggaaaaac aataggtcct taaataatat
     1681 tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct attctatatg
     1741 aaaagccggt tccggcgctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg
     1801 tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct
     1861 gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg gttaataact
     1921 tcgtataatg tatgctatac gaagttatcc cgggtaccga gctcgaattc gtaacttaca
     1981 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     2041 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     2101 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     2161 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     2221 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     2281 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     2341 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     2401 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     2461 atgaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2521 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2581 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2641 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2701 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2761 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2821 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2881 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2941 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3001 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3061 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3121 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3181 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3241 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3301 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3361 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3421 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3481 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3541 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3601 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3661 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3721 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3781 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3841 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3901 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3961 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4021 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4081 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4141 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4201 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4261 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4321 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4381 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4441 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4501 ctcttccttt ttcaatattg gccgcaaatt aaagccttcg agcgtcccaa aaccttctca
     4561 agcaaggttt tcagtataat gttacatgcg tacacgcgtc tgtacagaaa aaaaagaaaa
     4621 atttgaaata taaataacgt tcttaatact aacataacta taaaaaaata aatagggacc
     4681 tagacttcag gttgtctaac tccttccttt tcggttagag cggatgtggg gggagggcgt
     4741 gaatgtaagc gtgacataac taattacatg actcgagtgc gcagatatca ctagtatatt
     4801 tgttgtaaaa agtagataat tacttccttg atgatctgta aaaaagagaa aaagaaagca
     4861 tctaagaact tgaaaaacta cgaattagaa aagaccaaat atgtatttct tgcattgacc
     4921 aatttatgca agtttatata tatgtaaatg taagtttcac gaggttctac taaactaaac
     4981 cacccccttg gttagaagaa aagagtgtgt gagaacaggc tgttgttgtc acacgattcg
     5041 gacaattctg tttgaaagag agagagtaac agtacgatcg aacgaacttt gctctggaga
     5101 tcacagtggg catcatagca tgtggtacta aaccctttcc cgccattcca gaaccttcga
     5161 ttgcttgtta caaaacctgt gagccgtcgc taggaccttg ttgtgtgacg aaattggaag
     5221 ctgcaatcaa taggaagaca ggaagtcgag cgtgtctggg ttttttcagt tttgttcttt
     5281 ttgcaaacaa atcacgagcg acggtaattt ctttctcgat aagaggccac gtgctttatg
     5341 agggtaacat caattcaaga aggagggaaa cacttccttt ttctggccct gataatagta
     5401 tgagggtgaa gccaaaataa aggattcgcg cccaaatcgg catctttaaa tgcaggtatg
     5461 cgatagttcc tcactctttc cttactcacg agtaattctt gcaaatgcct aatattattg
     5521 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     5641 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
