LOCUS       sequence-248559-        3833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for PCR amplification of XdCrtE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3833)
  AUTHORS   Milne N, Tramontin LRR, Borodina I
  TITLE     A teaching protocol demonstrating the use of EasyClone and 
            CRISPR/Cas9 for metabolic engineering of Saccharomyces cerevisiae 
            and Yarrowia lipolytica.
  JOURNAL   FEMS Yeast Res. 2019 Sep 26. pii: foz062. doi: 
            10.1093/femsyr/foz062.
  PUBMED    31556952
REFERENCE   2  (bases 1 to 3833)
  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"; journalName: "FEMS Yeast 
            Res. 2019 Sep 26. pii: foz062. doi: 10.1093/femsyr/foz062."
COMMENT     Sequence Label: pCfB8741
FEATURES             Location/Qualifiers
     source          1..3833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     367..384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    501..522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(537..542,543..560,561..567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    575..591
                     /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)."
     primer_bind     580..602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     599..615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     599..615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1832..1849)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1832..1848)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1863)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2057..2076)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2176..2198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2236..2254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2322..2426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2427..2495,2496..3287)
                     /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     complement(2645..2664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3458..3833,1..213)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
       61 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
      421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      601 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga
      661 tccccacgcg attcacagag ggatatcggc tagctttttc aggatcgctt ccaatttgac
      721 gttcttcccc gttcttcctt ccgaatctac gtctccaaga tccatccttg agttagcttc
      781 tgcgaactct ccttgaagcc ttcctagttc tctctcgagt gcacctgaca aggtgttgag
      841 gacttcctga gtatattcga atgagtgggt ttctgtgcgc atgtagttta cacagtggtg
      901 aaggatctca ggagaggtcg atttcttctg caacgtattg atgacgagtc tcgatgaggg
      961 gttggcatga atcgagtgga tagtgggaaa actgaatttc ccttctgtga ggtcctctgc
     1021 aaaattctta ttatgggcat actcagaaga ctgaaggttc atatagtcat ccctgatctg
     1081 gaagtatatt gagatcaagt tgacaagctg gacaaagtct atgtcacatt ctgactttgc
     1141 catcatcaat ctgaccgcta tacggaacaa acctcccgtc tttccaagaa ccattttcac
     1201 atattcctct tcgctaggac acgtcagact atctctccag aatagctcca ggccttgccc
     1261 tctatggagg gaaagtatct cgtctgtgat cactttatca agatacgtat ctttcgagaa
     1321 cggaatctgc gaattaggag ttgacgtgcc cccgttttca gacgaggccg aggaggagga
     1381 ggatgcagag gagacggatg attgaagcga agcagatgaa ggaggaatta caggcatggg
     1441 tatcggtgtt gggcgaagct tgaagatctc ttgataagcc agaaagtaga cgtagtttgc
     1501 agtgtttatt gtctgcggaa tcccgtaaat tagatgggcc acaggcgacc cacgcctgag
     1561 gaccgatgaa tcctccacat cgtccattaa taagctagcg gtatgtagca tgccaacaac
     1621 gttctggatg acctcgagat cctccttctt gacatccaac caatagttga aagcctcgat
     1681 gagttgtgat cgaatttctt ttccagggtt ctttcctagg tagtgatacg gttcaaggag
     1741 cacgatatca tcctgaggag taaactcgag tggaattgct gtgaggatgt tcgcgtaatc
     1801 catatgacag atgggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact
     1861 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     1921 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     1981 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2041 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     2101 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2161 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     2221 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2281 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2341 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2401 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2461 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2521 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2581 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2641 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2701 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2761 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2821 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2881 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2941 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3001 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3061 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3121 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3181 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3241 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3301 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3361 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3421 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3481 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3541 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3601 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3661 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3721 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3781 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg aga
//
