LOCUS       sequence-69710-e        4747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for PCR-based transplant of Saccharomyces 
            cerevisiae TEF1 promoter in yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4747)
  AUTHORS   Horecka J, Chu AM, Davis RW
  TITLE     IpO: plasmids and methods for simplified, PCR-based DNA transplant 
            in yeast.
  JOURNAL   Yeast. 2014 May;31(5):185-93. doi: 10.1002/yea.3006. Epub 2014 Mar 
            20.
  PUBMED    24604451
REFERENCE   2  (bases 1 to 4747)
  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.3006"; 
            journalName: "Yeast"; date: "2014-05"; volume: "31"; issue: "5"; 
            pages: "185-93"
COMMENT     Sequence Label: pJH104
FEATURES             Location/Qualifiers
     source          1..4747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        378..593
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             594..1397
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(1892..1912)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1892..1910)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1931..1947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1931..1947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1944..1966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1955..1971
                     /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(1979..1985,1986..2003,2004..2009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2024..2045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2162..2179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2333..2921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2413..2432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3092..3883,3884..3952))
                     /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     3715..3734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3953..4057)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4084..4539
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4171..4190)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4381..4402
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4665..4687
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4679..4696
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4680..4696
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4706..4725
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4706..4724
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctcgagcgta cgctgcaggt cgacgaatcc ttacatcaca cccaatcccc cacaagtgat
       61 cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc
      121 ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact aaattccccc
      181 tctttcttcc tctagggtgg cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga
      241 gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt
      301 tttcttgaaa attttttttt ttgatttttt tctctttcga tgacctccca ttgattctag
      361 aagagtgcac cacgcttttc aattcatcat ttttttttta ttcttttttt tgatttcggt
      421 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
      481 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
      541 tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa atcatgtcga
      601 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
      661 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
      721 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
      781 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
      841 agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat
      901 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
      961 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     1021 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     1081 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     1141 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     1201 gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg
     1261 tggtctctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     1321 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     1381 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     1441 ttagagcttc aatttaatta tatcagttat taccctgcgg tgtgaaagga tcccacttca
     1501 aaacacccaa gcacagcata ctaaattccc cctctttctt cctctagggt ggcgttaatt
     1561 acccgtacta aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga
     1621 aaaaggcaat aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt
     1681 tttctctttc gatgacctcc cattgatatt taagttaata aatggtcttc aatttctcaa
     1741 gtttcagttt cgtttttctt gttctattac aacttttttt acttcttgct cattagaaag
     1801 aaagcatagc aatctatcta agttttaatt acaaacgagc tcgaattcat cgatgacggc
     1861 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     1921 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1981 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2041 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2101 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2161 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2221 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2281 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2341 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2401 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2461 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2521 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2581 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2641 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2701 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2761 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2821 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2881 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2941 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3001 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3061 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3121 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3181 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3241 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3301 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3361 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3421 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3481 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3541 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3601 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3661 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3721 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3781 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3841 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3901 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3961 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4021 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4081 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4141 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4201 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4261 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4321 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4381 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4441 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4501 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     4561 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4621 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     4681 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     4741 gcccccc
//
