LOCUS       sequence-180204-        9272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tetR-mTurqoise2 cloned between BamHI and EcoRI of pAM2991.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9272)
  AUTHORS   Zheng XY, O'Shea EK
  TITLE     Cyanobacteria Maintain Constant Protein Concentration despite Genome
            Copy-Number Variation.
  JOURNAL   Cell Rep. 2017 Apr 18;19(3):497-504. doi: 
            10.1016/j.celrep.2017.03.067.
  PUBMED    28423314
REFERENCE   2  (bases 1 to 9272)
  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: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-18- 18"; volume: "19"; issue: "3"; pages: "497-504"
COMMENT     Sequence Label: EB2317
FEATURES             Location/Qualifiers
     source          1..9272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..621
                     /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(74..93)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(1366..1383)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1536..1622
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1714..1741
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2206..2997)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    4916..5056
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4971..4990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5071..5088)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5242..5830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5322..5341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7801..7878
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             7879..8961
                     /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(7898..7917)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(9193..9198,9199..9215,9216..9222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    9230..9246
                     /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     9235..9257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgtctagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
       61 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
      121 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
      181 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
      241 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
      301 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
      361 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
      421 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
      481 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
      541 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
      601 cttaaatgtg aaagtgggtc tgacatcctc gagttgatgg tttcgaaggg cgaagaactg
      661 tttacgggag tggtgccgat cctggttgag ttggatggcg acgtgaacgg acacaagttt
      721 agtgtctccg gcgaaggtga gggtgacgct acctacggta aactgacgct gaaattcatt
      781 tgtactaccg gtaaactccc ggttccatgg cctacgctgg ttacgaccct cagctggggt
      841 gtccaatgct tcgcccgcta cccggaccac atgaagcagc acgacttttt caaaagtgcc
      901 atgcctgagg ggtacgttca agaacgcacc attttcttta aggatgacgg caattacaaa
      961 acacgcgcag aggtgaaatt tgaaggtgat acgttggtca atcgaatcga actcaaaggc
     1021 attgacttta aagaggatgg caatatcctc ggccacaaac tggaatacaa ttacttcagc
     1081 gacaacgtct atatcacagc tgataagcag aaaaatggga ttaaagccaa ctttaagatc
     1141 cggcataaca ttgaagatgg cggcgtgcaa ctggcggatc attatcagca gaatactccc
     1201 atcggcgatg ggcccgtcct gttgccagat aaccattatc tgtctaccca aagcaagttg
     1261 tcgaaggatc ccaacgagaa gcgtgatcat atggtcctct tggagttcgt gactgcggca
     1321 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1381 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1441 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1501 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1561 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1621 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1681 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1741 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1801 atggcggccc agatccgcgg ccgccgatcc tctagtatgc ttgtaaaccg ttttgtgaaa
     1861 aaatttttaa aataaaaaag gggacctcta gggtccccaa ttaattagta atataatcta
     1921 ttaaaggtca ttcaaaaggt catccaccgg atcagcttag taaagccctc gctagatttt
     1981 aatgcggatg ttgcgattac ttcgccaact attgcgataa caagaaaaag ccagcctttc
     2041 atgatatatc tcccaatttg tgtagggctt attatgcacg cttaaaaata ataaaagcag
     2101 acttgacctg atagtttggc tgtgagcaat tatgtgctta gtgcatctaa cgcttgagtt
     2161 aagccgcgcc gcgaagcggc gtcggcttga acgaattgtt agacattatt tgccgactac
     2221 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     2281 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     2341 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     2401 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     2461 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     2521 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     2581 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     2641 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     2701 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     2761 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     2821 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     2881 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     2941 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatgat
     3001 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     3061 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     3121 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     3181 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     3241 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     3301 cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg
     3361 caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa
     3421 ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc cgtcgcggcg
     3481 cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga
     3541 gcatcgtttg ttcgcccagc ttctgtatgg aacgggcatg cggatcagtg agggtttgca
     3601 actgcgggtc aaggatctgg atttcgatca cggcacgatc atcgtgcggg agggcaaggg
     3661 ctccaaggat cgggccttga tgttacccga gagcttggca cccagcctgc gcgagcaggg
     3721 gaattgatcc ggtggatgac cttttgaatg acctttaata gattatatta ctaattaatt
     3781 ggggacccta gaggtcccct tttttatttt aaaaattttt tcacaaaacg gtttacaagc
     3841 ataaagctct agagtcgacc tgcaggcatg caagcttcga gtccctgctc gtcacgcttt
     3901 caggcaccgt gccagatatc gacgtggagt cgatcactgt gattggcgaa ggggaaggca
     3961 gcgctaccca aatcgctagc ttgctggaga agctgaaaca aaccacgggc attgatctgg
     4021 cgaaatccct accgggtcaa tccgactcgc ccgctgcgaa gtcctaagag atagcgatgt
     4081 gaccgcgatc gcttgtcaag aatcccagtg atcccgaacc ataggaaggc aagctcaatg
     4141 cttgcctcgt cttgaggact atctagatgt ctgtggaacg cacatttatt gccatcaagc
     4201 ccgatggcgt tcagcggggt ttggtcggta cgatcatcgg ccgctttgag caaaaaggct
     4261 tcaaactggt gggcctaaag cagctgaagc ccagtcgcga gctggccgaa cagcactatg
     4321 ctgtccaccg cgagcgcccc ttcttcaatg gcctcgtcga gttcatcacc tctgggccga
     4381 tcgtggcgat cgtcttggaa ggcgaaggcg ttgtggcggc tgctcgcaag ttgatcggcg
     4441 ctaccaatcc gctgacggca gaaccgggca ccatccgtgg tgattttggt gtcaatattg
     4501 gccgcaacat catccatggc tcggatgcaa tcgaaacagc acaacaggaa attgctctct
     4561 ggtttagccc agcagagcta agtgattgga cccccacgat tcaaccctgg ctgtacgaat
     4621 aaggtctgca ttccttcaga gagacattgc catgcccgtg ctgcgatcgc ccttccaagc
     4681 tgccttgccc cgctgtttcg ggctggcagc cctggcgttg gggctggcga ccgcttgcca
     4741 agaaagcagc gctccgccgg ctgccggatc gatcctgcct cgcgcgtttc ggtgatgacg
     4801 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4861 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     4921 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     4981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5041 aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5101 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5641 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6001 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6061 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6301 cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6541 tgtgactggt gagtcgccgg ggctggcagc ttagtcctgc gcaatctcta ctacatctgc
     6601 caacccagtg aaattttgat ctttgctggc agtagtcgcc gcagtagtga tggccgccga
     6661 gttggctatc gcttggtcaa gggcggcagc agcctgcggg tacctctgct ggaaaaagcg
     6721 ctccgcatgg atctgaccaa catgatcatt gagttgcgcg tttccaatgc cttctccaag
     6781 ggcggcattc ccctgactgt tgaaggcgtt gccaatatca agattgctgg ggaagaaccg
     6841 accatccaca acgcgatcga gcggctgctt ggcaaaaacc gtaaggaaat cgagcaaatt
     6901 gccaaggaga ccctcgaagg caacttgcgt ggtgttttag ccagcctcac gccggagcag
     6961 atcaacgagg acaaaattgc ctttgccaaa agtctgctgg aagaggcgga ggatgacctt
     7021 gagcagctgg gtctagtcct cgatacgctg caagtccaga acatttccga tgaggtcggt
     7081 tatctctcgg ctagtggacg caagcagcgg gctgatctgc agcgagatgc ccgaattgct
     7141 gaagccgatg cccaggctgc ctctgcgatc caaacggccg aaaatgacaa gatcacggcc
     7201 ctgcgtcgga tcgatcgcga tgtagcgatc gcccaagccg aggccgagcg ccggattcag
     7261 gatgcgttga cgcggcgcga agcggtggtg gccgaagctg aagcggacat tgctaccgaa
     7321 gtcgctcgta gccaagcaga actccctgtg cagcaggagc ggatcaaaca ggtgcagcag
     7381 caacttcaag ccgatgtgat cgccccagct gaggcagctt gtaaacgggc gatcgcggaa
     7441 gcgcgggggg ccgccgcccg tatcgtcgaa gatggaaaag ctcaagcgga agggacccaa
     7501 cggctggcgg aggcttggca gaccgctggt gctaatgccc gcgacatctt cctgctccag
     7561 aagctcgaaa ttcgagctcg gtacccgggg atctgggccg cgggtcatgg ctgcgccccg
     7621 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     7681 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     7741 gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg catttacgtt
     7801 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     7861 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     7921 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     7981 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     8041 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     8101 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     8161 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     8221 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     8281 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     8341 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     8401 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     8461 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     8521 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     8581 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     8641 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     8701 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     8761 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     8821 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     8881 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     8941 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg aattgatctg
     9001 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
     9061 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
     9121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     9181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
     9241 taacaatttc acacaggaaa cagaccatgg aa
//
