LOCUS       sequence-12701-e        6551 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 6551)
  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 6551)
  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: pXP222
FEATURES             Location/Qualifiers
     source          1..6551
                     /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(439..472)
                     /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(1914..1947)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1975..3317
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2307..2354
                     /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"
     terminator      complement(5379..5626)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5592..5610
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     6524..6542
                     /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 atcccgggat aacttcgtat agcatacatt atacgaagtt attaaccatt
      481 atttttttcc tcaacataac gagaacacac aggggcgcta tcgcacagaa tcaaattcga
      541 tgactggaaa ttttttgtta atttcagagg tcgcctgacg catatacctt tttcaactga
      601 aaaattggga gaaaaaggaa aggtgagagc gccggaaccg gcttttcata tagaatagag
      661 aagcgttcat gactaaatgc ttgcatcaca atacttgaag ttgacaatat tatttaagga
      721 cctattgttt ttccaatagg tggttagcaa tcgtcttact ttctaacttt tcttaccttt
      781 tacatttcag caatatatat atatatttca aggatatacc attctaatgt ctgcccctaa
      841 gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa
      901 ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt
      961 aattggtggt gctgctatcg aagctacagg tgtcccactt ccagatgagg cgctggaagc
     1021 ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtacagg
     1081 tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa
     1141 cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca
     1201 atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg
     1261 taagagaaag gaagatgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc
     1321 agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt
     1381 gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac
     1441 tgtggaggaa accatcaaga acgaatttcc tacattgaag gttcaacatc aattgattga
     1501 ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag
     1561 caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt
     1621 gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga
     1681 accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat
     1741 cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat
     1801 tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc
     1861 caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccataactt
     1921 cgtatagcat acattatacg aagttatccc ggggtaccga gctcgaattc gtatgatcca
     1981 atatcaaagg aaatgatagc attgaaggat gagactaatc caattgagga gtggcagcat
     2041 atagaacagc taaagggtag tgctgaagga agcatacgat accccgcatg gaatgggata
     2101 atatcacagg aggtactaga ctacctttca tcctacataa atagacgcat ataagtacgc
     2161 atttaagcat aaacacgcac tatgccgttc ttctcatgta tatatatata caggcaacac
     2221 gcagatatag gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt tgcattttcg
     2281 gaagcgctcg ttttcggaaa cgctttgaag ttcctattcc gaagttccta ttctctagaa
     2341 agtataggaa cttcagagcg cttttgaaaa ccaaaagcgc tctgaagacg cactttcaaa
     2401 aaaccaaaaa cgcaccggac tgtaacgagc tactaaaata ttgcgaatac cgcttccaca
     2461 aacattgctc aaaagtatct ctttgctata tatctctgtg ctatatccct atataaccta
     2521 cccatccacc tttcgctcct tgaacttgca tctaaactcg acctctacat tttttatgtt
     2581 tatctctagt attactcttt agacaaaaaa attgtagtaa gaactattca tagagtgaat
     2641 cgaaaacaat acgaaaatgt aaacatttcc tatacgtagt atatagagac aaaatagaag
     2701 aaaccgttca taattttctg accaatgaag aatcatcaac gctatcactt tctgttcaca
     2761 aagtatgcgc aatccacatc ggtatagaat ataatcgggg atgcctttat cttgaaaaaa
     2821 tgcacccgca gcttcgctag taatcagtaa acgcgggaag tggagtcagg ctttttttat
     2881 ggaagagaaa atagacacca aagtagcctt cttctaacct taacggacct acagtgcaaa
     2941 aagttatcaa gagactgcat tatagagcgc acaaaggaga aaaaaagtaa tctaagatgc
     3001 tttgttagaa aaatagcgct ctcgggatgc atttttgtag aacaaaaaag aagtatagat
     3061 tctttgttgg taaaatagcg ctctcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     3121 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttttacaa aaatgaagca
     3181 cagattcttc gttggtaaaa tagcgctttc gcgttgcatt tctgttctgt aaaaatgcag
     3241 ctcagattct ttgtttgaaa aattagcgct ctcgcgttgc atttttgttc tacaaaatga
     3301 agcacagatg cttcgttgaa 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 tattggccgc aaattaaagc cttcgagcgt
     5401 cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac gcgtctgtac
     5461 agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat aactataaaa
     5521 aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt tagagcggat
     5581 gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgactcg agtgcgcaga
     5641 tatcactagt atatttgttg taaaaagtag ataattactt ccttgatgat ctgtaaaaaa
     5701 gagaaaaaga aagcatctaa gaacttgaaa aactacgaat tagaaaagac caaatatgta
     5761 tttcttgcat tgaccaattt atgcaagttt atatatatgt aaatgtaagt ttcacgaggt
     5821 tctactaaac taaaccaccc ccttggttag aagaaaagag tgtgtgagaa caggctgttg
     5881 ttgtcacacg attcggacaa ttctgtttga aagagagaga gtaacagtac gatcgaacga
     5941 actttgctct ggagatcaca gtgggcatca tagcatgtgg tactaaaccc tttcccgcca
     6001 ttccagaacc ttcgattgct tgttacaaaa cctgtgagcc gtcgctagga ccttgttgtg
     6061 tgacgaaatt ggaagctgca atcaatagga agacaggaag tcgagcgtgt ctgggttttt
     6121 tcagttttgt tctttttgca aacaaatcac gagcgacggt aatttctttc tcgataagag
     6181 gccacgtgct ttatgagggt aacatcaatt caagaaggag ggaaacactt cctttttctg
     6241 gccctgataa tagtatgagg gtgaagccaa aataaaggat tcgcgcccaa atcggcatct
     6301 ttaaatgcag gtatgcgata gttcctcact ctttccttac tcacgagtaa ttcttgcaaa
     6361 tgcctaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6481 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     6541 gccctttcgt c
//
