LOCUS       sequence-12713-e        6301 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 6301)
  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 6301)
  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: pXP422
FEATURES             Location/Qualifiers
     source          1..6301
                     /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)."
     rep_origin      1971..3313
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2934..2981)
                     /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(3333..3349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3333..3349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3346..3368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3357..3373
                     /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(3381..3387,3388..3405,3406..3411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3426..3447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3564..3581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3735..4323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3815..3834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4494..5285,5286..5354))
                     /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     5117..5136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        5413..5826
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5864..6111
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5880..5898)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6274..6292
                     /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 aacgaagcat
     1981 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     2041 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa
     2101 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2161 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca
     2221 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc
     2281 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg
     2341 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt
     2401 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc
     2461 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg
     2521 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga
     2581 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt
     2641 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact
     2701 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt
     2761 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag
     2821 caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt
     2881 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc
     2941 ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga
     3001 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta
     3061 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg
     3121 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct
     3181 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct
     3241 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct
     3301 ttgatattgg atcatacgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3361 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3421 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3541 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3601 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3661 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3721 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3781 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3841 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3901 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3961 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4021 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4081 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4141 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4201 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4261 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4321 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4381 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4441 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4501 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4561 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4621 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4681 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4741 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4801 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4861 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4921 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4981 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5041 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5101 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5161 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5221 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5281 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5341 tgttgaatac tcatactctt cctttttcaa tattaccgcg aatccttaca tcacacccaa
     5401 tcccccacaa gtgatccccc acacaccata gcttcaaaat gtttctactc cttttttact
     5461 cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc
     5521 atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt
     5581 ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt
     5641 tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc
     5701 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
     5761 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
     5821 tctaagtttt aattacaaaa ctagtgatat ctgcgcactc gagtcatgta attagttatg
     5881 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
     5941 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
     6001 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
     6061 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccaatat
     6121 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6181 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     6241 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     6301 c
//
