LOCUS       sequence-12707-e        5446 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 5446)
  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 5446)
  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: pXP322
FEATURES             Location/Qualifiers
     source          1..5446
                     /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)."
     protein_bind    complement(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"
     promoter        4558..4971
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5009..5256
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5025..5043)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5419..5437
                     /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 tattaaccat
      481 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
      541 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
      601 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
      661 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
      721 acctattgtt tttccaatag gtggttagca atcgtcttac tttctaactt ttcttacctt
      781 ttacatttca gcaatatata tatatatttc aaggatatac cattctaatg tctgccccta
      841 agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta
      901 aggttcttaa agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt
      961 taattggtgg tgctgctatc gaagctacag gtgtcccact tccagatgag gcgctggaag
     1021 cctccaagaa ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtacag
     1081 gtagtgttag acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca
     1141 acttaagacc atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac
     1201 aatttgctaa aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg
     1261 gtaagagaaa ggaagatgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc
     1321 cagaagtgca aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat
     1381 tgcctatttg gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa
     1441 ctgtggagga aaccatcaag aacgaatttc ctacattgaa ggttcaacat caattgattg
     1501 attctgccgc catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca
     1561 gcaacatgtt tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt
     1621 tgttgccatc tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg
     1681 aaccatgcca cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta
     1741 tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca
     1801 ttgaagatgc agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt
     1861 ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccataact
     1921 tcgtatagca tacattatac gaagttatcc cgggtaccga gctcgaattc atcacgtgct
     1981 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     2041 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     2101 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     2161 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     2221 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     2281 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     2341 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     2401 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     2461 acgaattcgt 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 ttcaatatta ccgcgaatcc ttacatcaca cccaatcccc cacaagtgat
     4561 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
     4621 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc
     4681 tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
     4741 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
     4801 tttcttgaaa attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta
     4861 agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa
     4921 ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gttttaatta
     4981 caaaactagt gatatctgcg cactcgagtc atgtaattag ttatgtcacg cttacattca
     5041 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     5101 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     5161 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     5221 gagaaggttt tgggacgctc gaaggcttta atttgcggcc aatattattg aagcatttat
     5281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5341 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5401 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
