LOCUS       sequence-162365-        5736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pACSA_J23113_tetR PEZ4_gfp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5736)
  AUTHORS   Zess EK, Begemann MB, Pfleger BF
  TITLE     Construction of new synthetic biology tools for the control of gene 
            expression in the cyanobacterium Synechococcus sp. strain PCC 7002.
  JOURNAL   Biotechnol Bioeng. 2016 Feb;113(2):424-32. doi: 10.1002/bit.25713. 
            Epub 2015 Sep 3.
  PUBMED    26192329
REFERENCE   2  (bases 1 to 5736)
  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/bit.25713";
            journalName: "Biotechnol Bioeng"; date: "2016-02"; volume: "113"; 
            issue: "2"; pages: "424-32"
COMMENT     Sequence Label: pEZ12
FEATURES             Location/Qualifiers
     source          1..5736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      474..1062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     963..982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1216..1233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1350..1371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1386..1391,1392..1409,1410..1416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1424..1440
                     /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     1429..1451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1448..1464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1448..1464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1482..1500
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1482..1499
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2060..2087)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2103..2174)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2222..2254)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     CDS             complement(2255..2875)
                     /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     2783..2802
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             2882..2893
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(2902..2936)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     primer_bind     complement(3793..3811)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3807..3854
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4358..4377)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4359..4377)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4384..4401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4384..4400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4393..4415)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4541..4996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4628..4647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4838..4859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5074..5178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5179..5247,5248..5736,1..303)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPEMNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5397..5416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
       61 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      121 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      181 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      241 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      301 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      361 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      421 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      481 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      541 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      601 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      661 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      721 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      781 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      841 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      901 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      961 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1021 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1081 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1141 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1201 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     1261 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     1321 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     1381 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     1441 tttcacacag gaaacagcta tgaccatgat tacgccaagc tatttaggtg acactataga
     1501 atactcaagc tatgaggtga ctgccgcact caatgttcaa ccagatctag tgattatttc
     1561 ggggttagca gccgatgcgg gcaacctggt gaagcaactg cgagaattag gttacaacgg
     1621 cattattgtt ggggggaacg gcttaaatac ttctaatatt ttccccgtct gccaagcaaa
     1681 atgtgatggg gtgttggtgg cccaagccta cagtgccgag ttagataatg agattaaccg
     1741 cgcgtttcgg gacgcctatt ttcaacaaaa ccaaaaagag ccgccccaat ttagtgccca
     1801 ggcttttacg gcgatccaag tttttgttga agccctcagc agcctcgatg aaaaaacgcc
     1861 cttagaaact cttgctctac cggacttgcg acgacaactg cgggacgaaa tttttgcagg
     1921 tacctacgtc acgcctttgg gtgaaatttc cttcacagag gaaggggaaa ttgtccagaa
     1981 ggaatttttt gtggcccaaa ttgaaatgga tgaatcgggt caacaggggc gtttcgcctt
     2041 cattgaaacg aactataaac gcagaaaggc ccacccgaag gtgagccagt gtgactctag
     2101 tagagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     2161 tttcgtttta tttgatgcct ggctctagta gtgatctaca ctagcactat cagtgttatt
     2221 aagctactaa agcgtagttt tcgtcgtttg cagcggaccc actttcacat ttaagttgtt
     2281 tttctaatcc gcatatgatc aattcaaggc cgaataagaa ggctggctct gcaccttggt
     2341 gatcaaataa ttcgatagct tgtcgtaata atggcggcat actatcagta gtaggtgttt
     2401 ccctttcttc tttagcgact tgatgctctt gatcttccaa tacgcaacct aaagtaaaat
     2461 gccccacagc gctgagtgca tataatgcat tctctagtga aaaaccttgt tggcataaaa
     2521 aggctaattg attttcgaga gtttcatact gtttttctgt aggccgtgta cctaaatgta
     2581 cttttgctcc atcgcgatga cttagtaaag cacatctaaa acttttagcg ttattacgta
     2641 aaaaatcttg ccagctttcc ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca
     2701 tctcaatggc taaggcgtcg agcaaagccc gcttattttt tacatgccaa tacaatgtag
     2761 gctgctctac acctagcttc tgggcgagtt tacgggttgt taaaccttcg attccgacct
     2821 cattaagcag ctctaatgcg ctgttaatca ctttactttt atctaatctg gacatctagt
     2881 atttctcctc tttctctagt agctagcata atccctagga ctgagctagc catcaggtta
     2941 acaaaaaagc aggaatatcc ctatcagtga tagacagaca tccctatcag tgatagatat
     3001 aatgttaact gtgggattgc aaaaggagct cccaactcat aaagtcaagt aggagattaa
     3061 tcatatgtcg gtcatcaagc ccgaaatgaa aattaaactg cgcatggagg gcgctgtgaa
     3121 cgggcacaag ttcgtgatcg aaggtgaagg cattggtaag ccgtatgaag gtactcaaac
     3181 cctggactta acggttaaag aaggtgcccc gttgccattt agctacgaca tcctgacacc
     3241 cgcttttcag tatggcaatc gcgcgttcac caagtatccg aaagatattc cggattattt
     3301 caagcaggca ttcccggaag gatactcatg ggaacgctcg atgacctacg aagaccaggg
     3361 catttgcatt gccacaagcg acatcacaat ggagggtgat tgcttttttt acaaaattcg
     3421 cttcgacggg acaaacttcc cgccgaatgg cccagtgatg cagaaaaaga cgcttaaatg
     3481 ggaaccgtcg acagagaaaa tgtacgttcg cgatggcgtt ctgaaaggcg atgttaatat
     3541 ggcgctgctt ctcgaagggg gaggccatta ccgttgtgat ttcaaaacca catacaaagc
     3601 aaaaaaagat gttcgtttgc cggacgcgca tgaagtcgat caccgcattg agatcctgtc
     3661 acatgataaa gattacaata aggtccgcct ttatgagcac gctgaggccc gctattctat
     3721 gctgcccagc caagctaaat aaggatccgg ctgctaacaa agcccgaaag gaagctgagt
     3781 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3841 tgacgggttt tttgtctaga gcgctcaagt tgatcgaatg tatgccccgg atatgaccta
     3901 gggagattct tctaaagcga gcagggtttg ccaagtggcg atcgcctcag ggaaccataa
     3961 ccaactttgt tcgaggcggc gggggttgag ctgggaacga tcgagatcta gggcgatcgc
     4021 ccggagtttt tcggctttcc ctacaagaat ttcctggtca acgtcggtgg gttgcctttg
     4081 ggccaaatcg tacatactaa tcgccaggcc cgcataaatc tccgcctgga gggctggggg
     4141 gaaatcctgg gtcgttgccg ccgctaaggc ttcataccaa cgatcattgg cggcttcgta
     4201 atctgcattg aggtaatgga caaaacccag ggccatcaaa atttcggggt cgttgggctg
     4261 ttgggctagg gccatgtccc aattgtcttg caccgtggcg atcgccgttg cttgggttgg
     4321 cgatggtggg acggcagcca aagttgttca gcatgcgccc tatagtgagt cgtattacaa
     4381 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     4441 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     4501 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca
     4561 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4621 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4681 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4741 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4801 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4861 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4921 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4981 ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5041 accgcatcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5101 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5161 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5221 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5281 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5341 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5401 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5461 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5521 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5581 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5641 gatcatgtaa ctcgccttga tcgttgggaa ccggagatga atgaagccat accaaacgac
     5701 gagcgtgaca ccacgatgcc tgtagcaatg gcaaca
//
