LOCUS       sequence-12708-e        7587 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 7587)
  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 7587)
  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: pXP412
FEATURES             Location/Qualifiers
     source          1..7587
                     /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(3201..3234)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      3261..4603
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    3593..3640
                     /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(4619..4635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4619..4635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4632..4654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4643..4659
                     /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(4667..4673,4674..4691,4692..4697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4712..4733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4850..4867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5021..5609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5101..5120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5780..6571,6572..6640))
                     /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     6403..6422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        6699..7112
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      7150..7397
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7166..7184)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     7560..7578
                     /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 tatgggccca
      481 ttatgaatac gcacctctat gtatttccga gatacgatta ctccagttcc tcttacaaga
      541 aatgcataaa aatagttaca attaattaga acaagaactt atttagaaca cgttcacact
      601 gagtaagaac tcttgtccct tattagcctt gatagtgctg aaaaaaagaa aaaaaacaaa
      661 aaaaagaaat aaaataacgg caaacagcaa aggccacaga accgtattca tgttacttct
      721 gcaatatcaa tcacttactg gcaagtgcgt ataaattaaa cctatttctt tatcatcata
      781 tttacttata tctttaacag attccaaacc ctaaagtgtc cgaattttca atagggcgaa
      841 cttgaagaat aaccaaggtc aataatatat cttttagtat aaccctgaaa tttgccctat
      901 agaaatctag ggtttctgtg tggtttccgg gtgagtcata cggctttttt gaatttcttt
      961 ttttgcagtt gtctctatca atgaaaattt cgaggaagac gataaggtta agataagtag
     1021 ataagagaat gatacgagat aaagcacaaa ttagcagaaa gaagagtggt tgcgaacaga
     1081 gtaaaccgaa tcagggaatc cctttttgca aaaacatcaa ttatcctttt ctttttttta
     1141 cgtatatatc tggaacagaa atatataagt tactattata cttatagttg gatccagttt
     1201 ttaatctgtc gtcaatcgaa agtttatttc agagttcttc agacttctta actcctgtaa
     1261 aaacaaaaaa aaaaaaaggc atagcaatga caaattcaaa agaagacgcc gacatagagg
     1321 agaagcatat gtacaatgag ccggtcacaa ccctctttca cgacgttgaa gcttcacaaa
     1381 cacaccacag acgtgggtca ataccattga aagatgagaa aagtaaagaa ttgtatccat
     1441 tgcgctcttt cccgacgaga gtaaatggcg aggatacgtt ctctatggag gatggcatag
     1501 gtgatgaaga tgaaggagaa gtacagaacg ctgaagtgaa gagagagctt aagcaaagac
     1561 atattggtat gattgccctt ggtggtacta ttggtacagg tcttttcatt ggtttatcca
     1621 cacctctgac caacgccggc ccagtgggcg ctcttatatc atatttattt atgggttctt
     1681 tggcatattc tgtcacgcag tccttgggtg aaatggctac attcatccct gttacatcct
     1741 ctttcacagt tttctcacaa agattccttt ctccagcatt tggtgcggcc aatggttaca
     1801 tgtattggtt ttcttgggca atcacttttg ccctggaact tagtgtagtt ggccaagtca
     1861 ttcaattttg gacgtacaaa gttccactgg cggcatggat tagtattttt tgggtaatta
     1921 tcacaataat gaacttgttc cctgtcaaat attacggtga attcgagttc tgggtcgctt
     1981 ccatcaaagt tttagccatt atcgggtttc taatatactg tttttgtatg gtttgtggtg
     2041 ctggggttac cggcccagtt ggattccgtt attggagaaa cccaggtgcc tggggtccag
     2101 gtataatatc taaggataaa aacgaaggga ggttcttagg ttgggtttcc tctttgatta
     2161 acgctgcctt cacatttcaa ggtactgaac tcgttggtat cactgctggt gaagctgcaa
     2221 accccagaaa atccgttcca agagccatca aaaaagttgt tttccgtatc ttaaccttct
     2281 acattggctc tctattattc attggacttt tagttccata caatgaccct aaactaacac
     2341 aatctacttc ctacgtttct acttctccct ttattattgc tattgagaac tctggtacaa
     2401 aggttttgcc acatatcttc aacgctgtta tcttaacaac cattatttct gccgcaaatt
     2461 caaatattta cgttggttcc cgtattttat ttggtctatc aaagaacaag ttggctccta
     2521 aattcctgtc aaggaccacc aaaggtggtg ttccatacat tgcagttttc gttactgctg
     2581 catttggcgc tttggcttac atggagacat ctactggtgg tgacaaagtt ttcgaatggc
     2641 tattaaatat cactggtgtt gcaggctttt ttgcatggtt atttatctca atctcgcaca
     2701 tcagatttat gcaagctttg aaataccgtg gcatctctcg tgacgagtta ccatttaaag
     2761 ctaaattaat gcccggcttg gcttattatg cggccacatt tatgacgatc attatcatta
     2821 ttcaaggttt cacggctttt gcaccaaaat tcaatggtgt tagctttgct gccgcctata
     2881 tctctatttt cctgttctta gctgtttgga tcttatttca atgcatattc agatgcagat
     2941 ttatttggaa gattggagat gtcgacatcg attccgatag aagagacatt gaggcaattg
     3001 tatgggaaga tcatgaacca aagacttttt gggacaaatt ttggaatgtt gtagcataga
     3061 tatgacgttt tattaccttt gatcacattt ccacgccatt tcgcattctc accctcataa
     3121 gtcatacacc gaaaagaaag tttaagggat caatgagctt actataatct cagtatattt
     3181 atttttatcg atgattcacc ataacttcgt atagcataca ttatacgaag ttatcccggg
     3241 taccgagctc gaattcgtat gatccaatat caaaggaaat gatagcattg aaggatgaga
     3301 ctaatccaat tgaggagtgg cagcatatag aacagctaaa gggtagtgct gaaggaagca
     3361 tacgataccc cgcatggaat gggataatat cacaggaggt actagactac ctttcatcct
     3421 acataaatag acgcatataa gtacgcattt aagcataaac acgcactatg ccgttcttct
     3481 catgtatata tatatacagg caacacgcag atataggtgc gacgtgaaca gtgagctgta
     3541 tgtgcgcagc tcgcgttgca ttttcggaag cgctcgtttt cggaaacgct ttgaagttcc
     3601 tattccgaag ttcctattct ctagaaagta taggaacttc agagcgcttt tgaaaaccaa
     3661 aagcgctctg aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact
     3721 aaaatattgc gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc
     3781 tctgtgctat atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta
     3841 aactcgacct ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg
     3901 tagtaagaac tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata
     3961 cgtagtatat agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc
     4021 atcaacgcta tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa
     4081 tcggggatgc ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc
     4141 gggaagtgga gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc
     4201 taaccttaac ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa
     4261 aggagaaaaa aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt
     4321 ttgtagaaca aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat
     4381 ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg
     4441 catttttgtt ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt
     4501 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     4561 cgttgcattt ttgttctaca aaatgaagca cagatgcttc gttgaattcg taatcatggt
     4621 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4681 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4861 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4921 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4981 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5461 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5761 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5821 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5881 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5941 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6001 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6061 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6121 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6181 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6241 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6301 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6361 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6421 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6481 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6541 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6601 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6661 accgcgaatc cttacatcac acccaatccc ccacaagtga tcccccacac accatagctt
     6721 caaaatgttt ctactccttt tttactcttc cagattttct cggactccgc gcatcgccgt
     6781 accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc ctctagggtg
     6841 tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc gtttcttttt
     6901 cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa aatttttttt
     6961 tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa acggtcttca
     7021 atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta cttcttgctc
     7081 attagaaaga aagcatagca atctaatcta agttttaatt acaaaactag tgatatctgc
     7141 gcactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg
     7201 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta
     7261 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     7321 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct
     7381 cgaaggcttt aatttgcggc caatattatt gaagcattta tcagggttat tgtctcatga
     7441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7501 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7561 ataggcgtat cacgaggccc tttcgtc
//
