LOCUS       sequence-67878-e        4121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pJH136 carries a URA3 cassette flanked by PCR priming sites U2 and 
            D2. URA3 was derived from S288c and extends from 243 bp upstream of 
            the start codon to 79 bp downstream of the stop codon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4121)
  AUTHORS   Horecka J, Davis RW
  TITLE     The 50:50 method for PCR-based seamless genome editing in yeast.
  JOURNAL   Yeast. 2014 Mar;31(3):103-12. doi: 10.1002/yea.2992. Epub 2013 Dec 
            13.
  PUBMED    24639370
REFERENCE   2  (bases 1 to 4121)
  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.2992"; 
            journalName: "Yeast"; date: "2014-03"; volume: "31"; issue: "3"; 
            pages: "103-12"
COMMENT     Sequence Label: pJH136
FEATURES             Location/Qualifiers
     source          1..4121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..260
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             261..1064
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     primer_bind     complement(1203..1219)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1204..1220)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1244..1263)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1245..1263)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1273..1290)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1273..1289)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1282..1304)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1431..1886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1518..1537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1728..1749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1912..2016
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2017..2085,2086..2877)
                     /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(2235..2254)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3048..3636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3537..3556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3790..3807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3924..3945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3960..3965,3966..3983,3984..3990)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3998..4014
                     /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     4003..4025
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4022..4038
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4022..4038
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4057..4077
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4059..4077
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 cgtacgctgc aggtcgactg tggtttcagg gtccataaag cttttcaatt catctttttt
       61 ttttttgttc ttttttttga ttccggtttc tttgaaattt ttttgattcg gtaatctccg
      121 agcagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgtg
      181 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct
      241 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
      301 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
      361 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
      421 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
      481 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa gacagaaaat
      541 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
      601 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
      661 agcaggcggc ggaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
      721 catgcaaggg ctccctagct actggagaat atactaaggg tactgttgac attgcgaaga
      781 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
      841 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gacgcattgg
      901 gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt attattgttg
      961 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     1021 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     1081 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     1141 ccgagctcga attcatcgat gggcggatcc cccgggctgc aggaattcga tatcaagctt
     1201 atcgataccg tcgacctcga gggggggccc ggtacccaat tcgccctata gtgagtcgta
     1261 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     1321 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     1381 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     1441 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1501 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1561 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1621 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1681 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1741 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1801 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1861 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     1921 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1981 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2041 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2101 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2161 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2221 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2281 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2341 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2401 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2461 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2521 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2581 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2641 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2701 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2761 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2821 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2881 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2941 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3001 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3061 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3181 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3721 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3781 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3841 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3901 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     3961 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     4021 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     4081 caaaagctgg agctccaccg cggtggcggc cgctctagcc c
//
