LOCUS       p2B58                   7098 bp    DNA     circular UNA 07-JUL-2021
DEFINITION  Insertion of pTET4-eGFP-tADH1 by Gibson Assembly in pRS426 backbone
            digested with HindIII/EcoRI.
ACCESSION   urn.local...77-dfzniz3
VERSION     urn.local...77-dfzniz3
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     promoter        190..415
     misc_feature    417..1217
                     /note="Geneious type: marker"
     misc_feature    complement(1483..1789)
                     /note="Geneious type: origin of replication"
     misc_feature    complement(1810..1970)
                     /note="Geneious type: reporter"
     primer_bind     1975..1994
     promoter        1977..1995
     misc_feature    2016..2040
     primer_bind     2016..2143
     primer_bind     complement(2016..2143)
     misc_feature    2046..2118
                     /note="Geneious type: Editing History Insertion"
     protein_bind    2046..2064
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2046..2118
     protein_bind    2119..2137
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2119..2481
     misc_feature    2119..2143
     primer_bind     2119..2143
     protein_bind    2185..2203
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2256..2275)
     protein_bind    2269..2287
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2439..2458
     primer_bind     complement(2455..2481)
     misc_feature    2457..2498
                     /note="Geneious type: Editing History Insertion"
     misc_feature    2457..2481
     primer_bind     2457..2501
     CDS             2499..3215
     source          <2499..>3423
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(2732..2751)
     primer_bind     3084..3103
     misc_feature    3223..3235
     terminator      3236..3423
                     /gene="S. cerevisiae ADH1"
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3401..3453)
     misc_feature    3424..3453
                     /note="Geneious type: Editing History Insertion"
     misc_feature    3429..3453
     promoter        complement(3495..3514)
     primer_bind     complement(3496..3515)
     misc_feature    complement(3553..3580)
                     /note="Geneious type: regulatory sequence"
     promoter        complement(3584..3613)
     misc_feature    complement(3922..4541)
                     /note="Geneious type: origin of replication"
     misc_feature    complement(4696..5556)
                     /note="Geneious type: marker"
     promoter        complement(5598..5626)
     misc_feature    complement(5688..6848)
                     /note="Geneious type: origin of replication"
     gene            complement(6651..6698)
ORIGIN      
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1861 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1921 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1981 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     2041 agctttccct atcagtgata gagaccttct ctttggaact ttcagtaata cgcttaactg
     2101 ctcattgcta tattgaagtc cctatcagtg atagagatgc gtcctcgtct tcaccggtcg
     2161 cgttcctgaa acgcagatgt gccttcccta tcagtgatag agaaacaata aagattctac
     2221 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggtc cctatcagtg
     2281 atagagataa ttaatcagcg aagcgatgat ttttgatcta ttaacagata tataaatgga
     2341 aaagctgcat aaccacttta actaatactt tcaacatttt cagtttgtat tacttcttat
     2401 tcaaatgtca taaaagtatc aacaaaaaat tgttaatata cctctatact ttaacgtcaa
     2461 ggagaaaaaa ctataactag agtgctggtt taattaacat gtcaaaaggc gaggaactat
     2521 tcacaggtgt cgtaccaata ctggtagaat tggatggcga cgtaaacggg cataagtttt
     2581 ctgtgagtgg tgagggtgaa ggggacgcaa cttatggcaa acttacactt aagttcatct
     2641 gcaccacagg gaagctgcct gtcccatggc caactctagt tacaactctc acttacggag
     2701 tacaatgttt ttctcgatac ccagaccaca tgaaacaaca tgactttttc aaatctgcca
     2761 tgcctgaggg atatgtgcaa gagagaacta ttttctttaa agatgatggt aattacaaaa
     2821 ctcgtgctga agtgaagttt gaaggagata cacttgttaa tagaattgag ttaaagggta
     2881 ttgactttaa ggaagatggt aatatcttag gtcacaaatt ggaatacaac tacaattcac
     2941 ataacgttta catcatggca gataaacaga aaaacggcat aaaagttaat ttcaaaatta
     3001 gacataatat cgaagatggt tccgttcaac tcgctgatca ttatcaacaa aatacaccaa
     3061 tcggagatgg acctgttcta ttgccagaca accattactt atctacacag agcgccttgt
     3121 caaaagatcc taatgaaaag agagatcaca tggttttgtt agaatttgtc accgctgcgg
     3181 gtataacgtt gggcatggat gaattataca agtaaggcgc gccacttcta aataagcgaa
     3241 tttcttatga tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa
     3301 attttaaagt gactcttagg ttttaaaacg aaaattctta ttcttgagta actctttcct
     3361 gtaggtcagg ttgctttctc aggtatagta tgaggtcgct cttattgacc acacctctac
     3421 cgggaattcc tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag
     3481 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     3541 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag
     3601 cataaagtgt aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg
     3661 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3721 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3781 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3841 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3901 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3961 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4021 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4081 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4141 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4201 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4261 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4321 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     4381 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4441 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4501 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4561 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4621 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4681 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4801 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4981 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5521 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5581 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5641 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg
     5701 tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     5761 tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa
     5821 tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     5881 gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca
     5941 aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa
     6001 caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata
     6061 actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc
     6121 ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg
     6181 tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat
     6241 actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg
     6301 gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt
     6361 ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag taatactaga
     6421 gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga
     6481 tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa
     6541 tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg
     6601 gttttttgaa agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta
     6661 tactttctag agaataggaa cttcggaata ggaacttcaa agcgtttccg aaaacgagcg
     6721 cttccgaaaa tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc gcacctatat
     6781 ctgcgtgttg cctgtatata tatatacatg agaagaacgg catagtgcgt gtttatgctt
     6841 aaatgcgtac ttatatgcgt ctatttatgt aggatgaaag gtagtctagt acctcctgtg
     6901 atattatccc attccatgcg gggtatcgta tgcttccttc agcactaccc tttagctgtt
     6961 ctatatgctg ccactcctca attggattag tctcatcctt caatgctatc atttcctttg
     7021 atattggatc atactaagaa accattatta tcatgacatt aacctataaa aataggcgta
     7081 tcacgaggcc ctttcgtc
//
