LOCUS       sequence-444180-        6549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inactive cogfp under control of Ptet, cogfp under control of Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6549)
  AUTHORS   Ljiljana Mihajlovic, Bharat Ravi Iyengar, Florian Baier, Icvara 
            Barbier, Justyna Iwaszkiewicz, Vincent Zoete, Andreas Wagner, 
            Yolanda Schaerli
  TITLE     A direct experimental test of Ohno's hypothesis
  JOURNAL   eLife13:RP97216
REFERENCE   2  (bases 1 to 6549)
  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: 
            "eLife13:RP97216"
COMMENT     Sequence Label: pDUP1
FEATURES             Location/Qualifiers
     source          1..6549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             269..892
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(342..361)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1075..1094)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(1466..2011)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1546..1565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2100..2915)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2100..2915)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2823..2842
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2907..2926
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3052..3069
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3053..3069
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3089..3107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3114..3132
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             3147..3169
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3183..3200
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3909..3936
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4694..4711)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4724..4746
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4751..4770
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(join(4776..4782,4783..4798,4799..4804))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             4925..6007
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLXKRKTXXAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4944..4963)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttatcgaaaa ataaatatag caaataataa atcaaacaac gatttaacca aaagagaaaa
       61 agaatgttta gcgtgggcat gcgaaggaaa aagctcttgg gatatttcaa aaatattagg
      121 ttgcagtgag cgtactgtca ctttccattt aaccaatgcg caaatgaaac tcaatacaac
      181 aaaccgctgc caaagtattt ctaaagcaat tttaacagga gcaattgatt gcccatactt
      241 taaaaattaa ggatccagga ggaaaaaaat gtccagatta gataaaagta aagtgattaa
      301 cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc
      361 ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt
      421 gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt gccctttaga
      481 aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact
      541 aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga
      601 aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt
      661 atatgcactc agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca
      721 tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg
      781 acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga
      841 attgattata tgcggattag aaaaacaact taaatgtgaa agtgggtcct aataattggt
      901 aacgaatcag acaattgacg gctcgaggga gtagcatagg gtttgcagaa tccctgcttc
      961 gtccatttga caggcacatt atgcatcgat gataagctgt caaacatgag cagatcctct
     1021 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
     1081 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcaaatatt
     1141 ttatctgagg tagagaccgc ttcctcgctc actgactcgc tgcacgaggc agacctcagc
     1201 gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct
     1261 tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata
     1321 tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa
     1381 atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa
     1441 gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg
     1501 aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt
     1561 ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca
     1621 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
     1681 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
     1741 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
     1801 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
     1861 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
     1921 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
     1981 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc
     2041 tcagtggaac gaaaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacct
     2101 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     2161 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2221 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2281 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2341 gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag
     2401 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     2461 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     2521 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     2581 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     2641 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     2701 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     2761 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     2821 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     2881 ctggagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     2941 taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat gtacatcaga
     3001 gattttgaga cacaaggtct cccgctcgac gtacggtgga agaattcgag ttgtaaaacg
     3061 acggccagta ctccaccgtt ggcttttttc cctatcagtg atagagattg acatccctat
     3121 cagtgataga gataatgagc actaattttg tttaacttta agaaggagat atacatatgg
     3181 gccatcatca ccatcatcac agcattccgg aaaatagcgg tctgaccgaa gaaatgcctg
     3241 cacagatgaa tctggaaggt gttgttaatg gtcatgcctt tagcatggaa ggtattggtg
     3301 gtggtaatat tctgaccggt attcagaaac tggatattcg tgttattgaa ggtgatccgc
     3361 tgccgtttag ctttgatatt ctgagcgttg catttgccag cgcgaatcgt acctatacca
     3421 gctatccggc aaaaatcccg gattattttg ttcagagctt tccggaaggt tttacctttg
     3481 aacgtaccct gagctttgaa gatggtgcca ttgttaaagt ggaaagcgat attagcatcg
     3541 aggatggtaa atttgtgggc aaaatcaaat ataacggtga aggtttcccg gaagatggtc
     3601 cggttatgaa aaaagaagtt accaaactgg aaccgggcag cgaaagcatg tatgttagtg
     3661 atggcaccct ggttggtgaa gttgttctga gctataaaac ccagagcacc cattatacct
     3721 gtcacatgaa aaccatttat cgcagcaaaa aaccggttga aaacctgccg aaatttcatt
     3781 atgttcatca ccgcctggaa aaaaaaattg tggaagaggg ctattattac gagcagcatg
     3841 aaaccgcaat tgccaaaccg taataagagc tccaatcgct tggaccagct ttccctgcag
     3901 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag agagaatata
     3961 aaaagccaga ttattaatcc ggctttttta ttatttgtcg acctcattcg ctaatcgcca
     4021 cggtacctta ttacggtttg gcaattgcgg tttcatgctg ctcgtaataa tagccctctt
     4081 ccacaatttt tttttccagg cggtgatgaa cataatgaaa tttcggcagg ttttcaaccg
     4141 gttttttgct gcgataaatg gttttcatgt gacaggtata atgggtgctc tgggttttat
     4201 agctcagaac aacttcacca accagggtgc catcactaac atacatgctt tcgctgcccg
     4261 gttccagttt ggtaacttct tttttcataa ccggaccatc ttccgggaaa ccttcaccgt
     4321 tatatttgat tttgcccaca aatttaccat cctcgatgct aatatcgctt tccactttaa
     4381 caatggcacc atcttcaaag ctcagggtac gttcaaaggt aaaaccttcc ggaaagctct
     4441 gaacaaaata atccgggatt tttgccggat agctggtata ggtacgattg ccatactgaa
     4501 atgcaacgct cagaatatca aagctaaacg gcagcggatc accttcaata acacgaatat
     4561 ccagtttctg aataccggtc agaatattac caccaccaat accttccatg ctaaaggcat
     4621 gaccattaac aacaccttcc agattcatct gtgcaggcat ttcttcggtc agaccgctat
     4681 tttccggaat gctgtgatga tggtgatgat ggcccatggt atatctcctt cttaaagtta
     4741 aacaaaatta aattgtgagc gctcacaatt ccacacatta tacgagccga tgattaattg
     4801 tcaacagctc gtaaaagacg cagtgacggc gcggccgcaa tggcggcgcg ccatcgaatg
     4861 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     4921 gaatatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     4981 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     5041 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     5101 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     5161 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     5221 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     5281 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     5341 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     5401 tattttctcc catgaggacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     5461 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     5521 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     5581 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     5641 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     5701 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagatag
     5761 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     5821 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     5881 gccagtctca ctggkgaaaa gaaaaaccmc ccyggcgccc aayacgcaaa ccgcctctcc
     5941 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6001 gcagtgataa gatcctattc cagcgggtca cacaggaaag gcctcgatga aaaacataaa
     6061 tgccgacgac acatacagaa taattaataa aattaaagct tgtagaagca ataatgatat
     6121 taatcaatgc ttatctgata tgactaaaat ggtacattgt gaatattatt tactcgcgat
     6181 catttatcct cattctatgg ttaaatctga tatttcaatc ctagataatt accctaaaaa
     6241 atggaggcaa tattatgatg acgctaattt aataaaatat gatcctatag tagattattc
     6301 taactccaat cattcaccaa ttaattggaa tatatttgaa aacaatgctg taaataaaaa
     6361 atctccaaat gtaattaaag aagcgaaaac atcaggtctt atcactgggt ttagtttccc
     6421 tattcatacg gctaacaatg gcttcggaat gcttagtttt gcacattcag aaaaagacaa
     6481 ctatatagat agtttatttt tacatgcgtg tatgaacata ccattaattg ttccttctct
     6541 agttgataa
//
