LOCUS       sequence-162049-        5736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pACSA_J23113_tetR PEZ2_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: pEZ08
FEATURES             Location/Qualifiers
     source          1..5736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..83)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     274..295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        510..614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(615..683,684..1475)
                     /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(833..852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1646..2234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2135..2154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2388..2405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2522..2543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2558..2563,2564..2581,2582..2588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2596..2612
                     /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     2601..2623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2620..2636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2620..2636
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2654..2672
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2654..2671
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(3232..3259)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3275..3346)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3394..3426)
                     /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(3427..4047)
                     /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     3955..3974
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             4054..4065
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(4074..4108)
                     /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(4965..4983)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4979..5026
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5530..5549)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5531..5549)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5556..5573)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5556..5572)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5565..5587)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(5713..5736,1..432)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
       61 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      181 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      301 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      421 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      481 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1081 atgtaactcg ccttgatcgt tgggaaccgg agatgaatga agccatacca aacgacgagc
     1141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2341 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2401 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2461 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2521 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     2581 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     2641 attacgccaa gctatttagg tgacactata gaatactcaa gctatgaggt gactgccgca
     2701 ctcaatgttc aaccagatct agtgattatt tcggggttag cagccgatgc gggcaacctg
     2761 gtgaagcaac tgcgagaatt aggttacaac ggcattattg ttggggggaa cggcttaaat
     2821 acttctaata ttttccccgt ctgccaagca aaatgtgatg gggtgttggt ggcccaagcc
     2881 tacagtgccg agttagataa tgagattaac cgcgcgtttc gggacgccta ttttcaacaa
     2941 aaccaaaaag agccgcccca atttagtgcc caggctttta cggcgatcca agtttttgtt
     3001 gaagccctca gcagcctcga tgaaaaaacg cccttagaaa ctcttgctct accggacttg
     3061 cgacgacaac tgcgggacga aatttttgca ggtacctacg tcacgccttt gggtgaaatt
     3121 tccttcacag aggaagggga aattgtccag aaggaatttt ttgtggccca aattgaaatg
     3181 gatgaatcgg gtcaacaggg gcgtttcgcc ttcattgaaa cgaactataa acgcagaaag
     3241 gcccacccga aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga
     3301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag
     3361 tagtgatcta cactagcact atcagtgtta ttaagctact aaagcgtagt tttcgtcgtt
     3421 tgcagcggac ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag
     3481 gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa
     3541 taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc
     3601 ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc
     3661 attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata
     3721 ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa
     3781 agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa
     3841 agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc
     3901 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag
     3961 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat
     4021 cactttactt ttatctaatc tggacatcta gtatttctcc tctttctcta gtagctagca
     4081 taatccctag gactgagcta gccatcaggt taacaaaaaa gcaggaatat ccctatcagt
     4141 gatagacaga catccctatc agtgatagat ataaagttaa ctgtgggatt gcaaaaggag
     4201 ctcccaactc ataaagtcaa gtaggagatt aatcatatgt cggtcatcaa gcccgaaatg
     4261 aaaattaaac tgcgcatgga gggcgctgtg aacgggcaca agttcgtgat cgaaggtgaa
     4321 ggcattggta agccgtatga aggtactcaa accctggact taacggttaa agaaggtgcc
     4381 ccgttgccat ttagctacga catcctgaca cccgcttttc agtatggcaa tcgcgcgttc
     4441 accaagtatc cgaaagatat tccggattat ttcaagcagg cattcccgga aggatactca
     4501 tgggaacgct cgatgaccta cgaagaccag ggcatttgca ttgccacaag cgacatcaca
     4561 atggagggtg attgcttttt ttacaaaatt cgcttcgacg ggacaaactt cccgccgaat
     4621 ggcccagtga tgcagaaaaa gacgcttaaa tgggaaccgt cgacagagaa aatgtacgtt
     4681 cgcgatggcg ttctgaaagg cgatgttaat atggcgctgc ttctcgaagg gggaggccat
     4741 taccgttgtg atttcaaaac cacatacaaa gcaaaaaaag atgttcgttt gccggacgcg
     4801 catgaagtcg atcaccgcat tgagatcctg tcacatgata aagattacaa taaggtccgc
     4861 ctttatgagc acgctgaggc ccgctattct atgctgccca gccaagctaa ataaggatcc
     4921 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     4981 agcataaccc cttggggcct ctaaacgggt cttgacgggt tttttgtcta gagcgctcaa
     5041 gttgatcgaa tgtatgcccc ggatatgacc tagggagatt cttctaaagc gagcagggtt
     5101 tgccaagtgg cgatcgcctc agggaaccat aaccaacttt gttcgaggcg gcgggggttg
     5161 agctgggaac gatcgagatc tagggcgatc gcccggagtt tttcggcttt ccctacaaga
     5221 atttcctggt caacgtcggt gggttgcctt tgggccaaat cgtacatact aatcgccagg
     5281 cccgcataaa tctccgcctg gagggctggg gggaaatcct gggtcgttgc cgccgctaag
     5341 gcttcatacc aacgatcatt ggcggcttcg taatctgcat tgaggtaatg gacaaaaccc
     5401 agggccatca aaatttcggg gtcgttgggc tgttgggcta gggccatgtc ccaattgtct
     5461 tgcaccgtgg cgatcgccgt tgcttgggtt ggcgatggtg ggacggcagc caaagttgtt
     5521 cagcatgcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     5581 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     5641 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     5701 aatggcgaat ggacgcgccc tgtagcggcg cattaa
//
