LOCUS       sequence-444179-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cogfp copy 1 under control of Ptet, cogfp copy 2 under control of 
            Ptac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5008)
  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 5008)
  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: pDUP2
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    34..53
                     /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(59..65,66..81,82..87))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             208..1290
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(227..246)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2101..2724
                     /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(2174..2193)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2907..2926)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      complement(3298..3843)
                     /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(3378..3397)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3932..4747)
                     /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(3932..4747)
                     /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     4655..4674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4739..4758
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4884..4901
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4885..4901
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4921..4939
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4946..4964
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             4979..5001
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 ggtatatctc cttcttaaag ttaaacaaaa ttaaattgtg agcgctcaca attccacaca
       61 ttatacgagc cgatgattaa ttgtcaacag ctcgtaaaag acgcagtgac ggcgcggccg
      121 caatggcggc gcgccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
      181 gaagagagtc aattcagggt ggtgaatatg aaaccagtaa cgttatacga tgtcgcagag
      241 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
      301 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
      361 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
      421 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
      481 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
      541 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
      601 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
      661 gaccagacac ccatcaacag tattattttc tcccatgagg acggtacgcg actgggcgtg
      721 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
      781 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
      841 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
      901 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
      961 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
     1021 ggatacgacg ataccgaaga tagctcatgt tatatcccgc cgttaaccac catcaaacag
     1081 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
     1141 gcggtgaagg gcaatcagct gttgccagtc tcactggtga aaagaaaaac caccctggcg
     1201 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     1261 caggtttccc gactggaaag cgggcagtga taagatccta ttccagcggg tcacacagga
     1321 aaggcctcga tgaaaaacat aaatgccgac gacacataca gaataattaa taaaattaaa
     1381 gcttgtagaa gcaataatga tattaatcaa tgcttatctg atatgactaa aatggtacat
     1441 tgtgaatatt atttactcgc gatcatttat cctcattcta tggttaaatc tgatatttca
     1501 atcctagata attaccctaa aaaatggagg caatattatg atgacgctaa tttaataaaa
     1561 tatgatccta tagtagatta ttctaactcc aatcattcac caattaattg gaatatattt
     1621 gaaaacaatg ctgtaaataa aaaatctcca aatgtaatta aagaagcgaa aacatcaggt
     1681 cttatcactg ggtttagttt ccctattcat acggctaaca atggcttcgg aatgcttagt
     1741 tttgcacatt cagaaaaaga caactatata gatagtttat ttttacatgc gtgtatgaac
     1801 ataccattaa ttgttccttc tctagttgat aattatcgaa aaataaatat agcaaataat
     1861 aaatcaaaca acgatttaac caaaagagaa aaagaatgtt tagcgtgggc atgcgaagga
     1921 aaaagctctt gggatatttc aaaaatatta ggttgcagtg agcgtactgt cactttccat
     1981 ttaaccaatg cgcaaatgaa actcaataca acaaaccgct gccaaagtat ttctaaagca
     2041 attttaacag gagcaattga ttgcccatac tttaaaaatt aaggatccag gaggaaaaaa
     2101 atgtccagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
     2161 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
     2221 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
     2281 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
     2341 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
     2401 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
     2461 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
     2521 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
     2581 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
     2641 ggtgcagagc cagccttctt attcggcctt gaattgatta tatgcggatt agaaaaacaa
     2701 cttaaatgtg aaagtgggtc ctaataattg gtaacgaatc agacaattga cggctcgagg
     2761 gagtagcata gggtttgcag aatccctgct tcgtccattt gacaggcaca ttatgcatcg
     2821 atgataagct gtcaaacatg agcagatcct ctacgccgga cgcatcgtgg ccggcatcac
     2881 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     2941 ggctcgccac ttcgggctca tgagcaaata ttttatctga ggtagagacc gcttcctcgc
     3001 tcactgactc gctgcacgag gcagacctca gcgctagcgg agtgtatact ggcttactat
     3061 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3121 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3181 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3241 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3301 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3361 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3421 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3481 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3541 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3601 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3661 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3721 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3781 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3841 caagaagatc atcttattaa ggggtctgac gctcagtgga acgaaaaatc aatctaaagt
     3901 atatatgagt aaacttggtc tgacagttac cttagaaaaa ctcatcgagc atcaaatgaa
     3961 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     4021 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     4081 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     4141 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     4201 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     4261 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     4321 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     4381 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     4441 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     4501 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     4561 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     4621 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     4681 aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat
     4741 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     4801 atatattttt atcttgtgca atgtacatca gagattttga gacacaaggt ctcccgctcg
     4861 acgtacggtg gaagaattcg agttgtaaaa cgacggccag tactccaccg ttggcttttt
     4921 tccctatcag tgatagagat tgacatccct atcagtgata gagataatga gcactaattt
     4981 tgtttaactt taagaaggag atatacat
//
