LOCUS       sequence-12702-e        6732 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 6732)
  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 6732)
  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: pXP312
FEATURES             Location/Qualifiers
     source          1..6732
                     /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    3201..3234
                     /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(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3777..3799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3788..3804
                     /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(3812..3818,3819..3836,3837..3842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3857..3878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3995..4012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4166..4754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4246..4265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4925..5716,5717..5785))
                     /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     5548..5567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        5844..6257
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      6295..6542
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6311..6329)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6705..6723
                     /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 tatggtgaat
      481 catcgataaa aataaatata ctgagattat agtaagctca ttgatccctt aaactttctt
      541 ttcggtgtat gacttatgag ggtgagaatg cgaaatggcg tggaaatgtg atcaaaggta
      601 ataaaacgtc atatctatgc tacaacattc caaaatttgt cccaaaaagt ctttggttca
      661 tgatcttccc atacaattgc ctcaatgtct cttctatcgg aatcgatgtc gacatctcca
      721 atcttccaaa taaatctgca tctgaatatg cattgaaata agatccaaac agctaagaac
      781 aggaaaatag agatataggc ggcagcaaag ctaacaccat tgaattttgg tgcaaaagcc
      841 gtgaaacctt gaataatgat aatgatcgtc ataaatgtgg ccgcataata agccaagccg
      901 ggcattaatt tagctttaaa tggtaactcg tcacgagaga tgccacggta tttcaaagct
      961 tgcataaatc tgatgtgcga gattgagata aataaccatg caaaaaagcc tgcaacacca
     1021 gtgatattta atagccattc gaaaactttg tcaccaccag tagatgtctc catgtaagcc
     1081 aaagcgccaa atgcagcagt aacgaaaact gcaatgtatg gaacaccacc tttggtggtc
     1141 cttgacagga atttaggagc caacttgttc tttgatagac caaataaaat acgggaacca
     1201 acgtaaatat ttgaatttgc ggcagaaata atggttgtta agataacagc gttgaagata
     1261 tgtggcaaaa cctttgtacc agagttctca atagcaataa taaagggaga agtagaaacg
     1321 taggaagtag attgtgttag tttagggtca ttgtatggaa ctaaaagtcc aatgaataat
     1381 agagagccaa tgtagaaggt taagatacgg aaaacaactt ttttgatggc tcttggaacg
     1441 gattttctgg ggtttgcagc ttcaccagca gtgataccaa cgagttcagt accttgaaat
     1501 gtgaaggcag cgttaatcaa agaggaaacc caacctaaga acctcccttc gtttttatcc
     1561 ttagatatta tacctggacc ccaggcacct gggtttctcc aataacggaa tccaactggg
     1621 ccggtaaccc cagcaccaca aaccatacaa aaacagtata ttagaaaccc gataatggct
     1681 aaaactttga tggaagcgac ccagaactcg aattcaccgt aatatttgac agggaacaag
     1741 ttcattattg tgataattac ccaaaaaata ctaatccatg ccgccagtgg aactttgtac
     1801 gtccaaaatt gaatgacttg gccaactaca ctaagttcca gggcaaaagt gattgcccaa
     1861 gaaaaccaat acatgtaacc attggccgca ccaaatgctg gagaaaggaa tctttgtgag
     1921 aaaactgtga aagaggatgt aacagggatg aatgtagcca tttcacccaa ggactgcgtg
     1981 acagaatatg ccaaagaacc cataaataaa tatgatataa gagcgcccac tgggccggcg
     2041 ttggtcagag gtgtggataa accaatgaaa agacctgtac caatagtacc accaagggca
     2101 atcataccaa tatgtctttg cttaagctct ctcttcactt cagcgttctg tacttctcct
     2161 tcatcttcat cacctatgcc atcctccata gagaacgtat cctcgccatt tactctcgtc
     2221 gggaaagagc gcaatggata caattcttta cttttctcat ctttcaatgg tattgaccca
     2281 cgtctgtggt gtgtttgtga agcttcaacg tcgtgaaaga gggttgtgac cggctcattg
     2341 tacatatgct tctcctctat gtcggcgtct tcttttgaat ttgtcattgc tatgcctttt
     2401 tttttttttg tttttacagg agttaagaag tctgaagaac tctgaaataa actttcgatt
     2461 gacgacagat taaaaactgg atccaactat aagtataata gtaacttata tatttctgtt
     2521 ccagatatat acgtaaaaaa aagaaaagga taattgatgt ttttgcaaaa agggattccc
     2581 tgattcggtt tactctgttc gcaaccactc ttctttctgc taatttgtgc tttatctcgt
     2641 atcattctct tatctactta tcttaacctt atcgtcttcc tcgaaatttt cattgataga
     2701 gacaactgca aaaaaagaaa ttcaaaaaag ccgtatgact cacccggaaa ccacacagaa
     2761 accctagatt tctatagggc aaatttcagg gttatactaa aagatatatt attgaccttg
     2821 gttattcttc aagttcgccc tattgaaaat tcggacactt tagggtttgg aatctgttaa
     2881 agatataagt aaatatgatg ataaagaaat aggtttaatt tatacgcact tgccagtaag
     2941 tgattgatat tgcagaagta acatgaatac ggttctgtgg cctttgctgt ttgccgttat
     3001 tttatttctt ttttttgttt tttttctttt tttcagcact atcaaggcta ataagggaca
     3061 agagttctta ctcagtgtga acgtgttcta aataagttct tgttctaatt aattgtaact
     3121 atttttatgc atttcttgta agaggaactg gagtaatcgt atctcggaaa tacatagagg
     3181 tgcgtattca taatgggccc ataacttcgt ataatgtatg ctatacgaag ttatcccggg
     3241 taccgagctc gaattcatca cgtgctataa aaataattat aatttaaatt ttttaatata
     3301 aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt
     3361 tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt atcttgctct
     3421 tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac
     3481 gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct atttaatctg
     3541 cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct
     3601 ttgtttattt ttttttcttc attccgtaac tcttctacct tctttattta ctttctaaaa
     3661 tccaaataca aaacataaaa ataaataaac acagagtaaa ttcccaaatt attccatcat
     3721 taaaagatac gaggcgcgtg taagttacga attcgtaatc atggtcatag ctgtttcctg
     3781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4861 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4921 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4981 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5041 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5101 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5161 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5221 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5281 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5341 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5401 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5461 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5521 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5581 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5641 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5701 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5761 cgacacggaa atgttgaata ctcatactct tcctttttca atattaccgc gaatccttac
     5821 atcacaccca atcccccaca agtgatcccc cacacaccat agcttcaaaa tgtttctact
     5881 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
     5941 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
     6001 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
     6061 caataaaaat ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc
     6121 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     6181 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
     6241 tagcaatcta atctaagttt taattacaaa actagtgata tctgcgcact cgagtcatgt
     6301 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     6361 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     6421 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     6481 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     6541 gcggccaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6601 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6661 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     6721 ggccctttcg tc
//
