LOCUS       sequence-69693-e        4605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template plasmid for PCR-based transplant of Ashbya gossypii TEF1 
            promoter in yeast. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4605)
  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 4605)
  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: pJH124
FEATURES             Location/Qualifiers
     source          1..4605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1035..1251
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(1760..1779)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1761..1779)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1789..1806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1789..1805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1798..1820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(1946..2401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2083..2104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2295..2314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2428..2532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2533..2601,2602..3393)
                     /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(2751..2770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3564..4152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4053..4072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4306..4323
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4440..4461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4476..4481,4482..4499,4500..4506)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4514..4530
                     /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     4519..4541
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4538..4554
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4538..4554
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4573..4593
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4575..4593
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
       61 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
      121 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
      181 agcggtttga agcaggcggc ggaagaagta acaaaggaac ctagaggcct tttgatgtta
      241 gcagaattgt catgcaaggg ctccctagct actggagaat atactaaggg tactgttgac
      301 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
      361 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
      421 gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
      481 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
      541 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
      601 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
      661 cagttattac ccggccgctc tagaccggtg catgcgctag ctgtgactgt cgcccgtaca
      721 tttagcccat acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg
      781 cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca
      841 cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt
      901 tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc
      961 acatcacatc cgaacataaa caaaaccgcg gtgtacagac gtcctcgagt gtggtttcag
     1021 ggtccataaa gcttttcaat tcatcttttt tttttttgtt cttttttttg attccggttt
     1081 ctttgaaatt tttttgattc ggtaatctcc gagcagaagg aagaacgaag gaaggagcac
     1141 agacttagat tggtatatat acgcatatgt ggtgttgaag aaacatgaaa ttgcccagta
     1201 ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac gaagataaat catgtcgaaa
     1261 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     1321 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     1381 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     1441 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     1501 tacaattttt tactcttcga agacagaaaa tttgctgaca ttggtaatac agtcaaattg
     1561 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     1621 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cggaagaagt aacaaaggaa
     1681 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctagc tactggagaa
     1741 tatactggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     1801 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1861 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1921 gttgcgcagc ctgaatggcg aatggaaatt gtaagcgtta atattttgtt aaaattcgcg
     1981 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2041 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2101 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2161 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2221 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2281 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2341 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2401 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2461 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2521 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2581 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2641 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2701 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2761 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2821 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2881 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2941 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3001 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3061 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3121 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3241 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3301 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3361 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4201 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4261 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4321 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4381 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4441 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4501 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4561 tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctg
//
