LOCUS       sequence-121994-        7198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cyclic nucleotide reporter for bacteria or plant cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7198)
  AUTHORS   Wheeler JI, Freihat L, Irving HR
  TITLE     A cyclic nucleotide sensitive promoter reporter system suitable for 
            bacteria and plant cells.
  JOURNAL   BMC Biotechnol. 2013 Nov 9;13:97. doi: 10.1186/1472-6750-13-97.
  PUBMED    24206622
REFERENCE   2  (bases 1 to 7198)
  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: "BMC 
            Biotechnol."; date: "2013-11-9"; volume: "13:97. doi"; pages: " 
            10.1186/1472-6750-13-97"
COMMENT     Sequence Label: pOPTXGARE
FEATURES             Location/Qualifiers
     source          1..7198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(309..328)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             320..1135
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(393..412)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1235..1823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1724..1743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1914..1938
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    1949..1973
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2571..2588)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3149..3173)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3178..4830
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3244..3262)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4985..5001)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4986..5002)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5026..5045)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5027..5045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5052..5069)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5052..5068)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5061..5083)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        5315..5494
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5528..6322
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5582..5601)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6192..6211
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6362..6613
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6637..6659
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      join(6792..7198,1..29)
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
ORIGIN
        1 agcccgctcg acggcgggct gttggatggg gatcgcctga atcgccccat catccagcca
       61 gaaagtgagg gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt
      121 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
      181 caactcagca aaagttcgat ttattcaaca aagccacgtt gtgtctcaaa atctctgatg
      241 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
      301 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ccaggccgcg
      361 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
      421 gcaatcaggt gcgacaatct atggattgta tgggaagccc gatgcgccag agttgtttct
      481 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
      541 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
      601 atggttactc accactgcga tcccagggaa aacagcattc caggtattag aagaatatcc
      661 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
      721 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
      781 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
      841 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
      901 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
      961 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1021 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1081 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     1141 attggttaat tggttgtaac catgaccaaa atcccttaac gtgagttttc ttccactgag
     1201 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1261 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1321 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1381 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1441 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1501 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1561 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1621 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1681 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1741 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1801 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1861 tttgctggcc ttttgctcac atgagatctc aaacaaacac atacagcgac ttagtttacc
     1921 cgccaatata tcctgtcaag gcctatcaac aagtttgtac aaaaaagcag gctgaaacgt
     1981 ttttcgttaa aaacttcacc gtaaattcca acaaatgtca aacaaaaaaa ttatcagaac
     2041 tatccaacaa attggtcata taattattca atggaaacgt ttccaataaa ttcgtgataa
     2101 atgcagattt ccaacgatca tattccaaca aatatatgtt ctattgaaaa tttattggaa
     2161 acatataatt cttacaaatt aaccgcatag atagctgttg gaattaagct tttttcttgt
     2221 agatcaaatt gaaaatattt ttcaccacat ccatcatttt cttgatttat tgtatataaa
     2281 tatgtacatt agatttctaa ttactttatc aacttttgtg aaatccataa aattttctta
     2341 aaattaaatt atgatgatat tgtaaacttg attagttctt ttaatgttaa gtttctacta
     2401 gtactgacgg aaagggttga actcacattg atgggaaggg aagatcgtca aagattgtca
     2461 tcataatcaa atcgcaacta tatatattca gccgtaaatg tcaagaaaac attgtagtaa
     2521 acaatagttt atgccccaga gccacaggcc aaagttgaaa ttttcttgta atgatgatga
     2581 tgatgatgca ttttaaacat attggacagt gggtccattt cttcccacta ctctaataca
     2641 ctaagaagtt tagtaaactt aattatttga tatatttaag ttttaaatta gttacacaat
     2701 ttattcaata gtattttgat atttttctaa taactaaaac cataatttta aaaatgaact
     2761 tctctataag aattcattgt ttctcttttt acacaaaagt tgatttaaga atttttttta
     2821 ctttttatta gaaagattta tatttattgg atattgatat tgaagtctag tttctaacaa
     2881 aaattcctaa caaagagagc ctaacaaaga gagcctaaca aagagagcct aacaaagaga
     2941 gcctaacaaa gagaatgtag aaatagttga ttaatattgg ataatatttt tctagtttct
     3001 ctttcatgat tcttttaaaa atgaacgaga cattataagt acctcctcgc tccccaccta
     3061 gttaatttaa ctttgtgcca actcctaaag ccaaacttat atcttccttt gtttgacata
     3121 atacaagaaa aaaagaaaag agctatggac ccagctttct tgtacaaagt ggttgccatg
     3181 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
     3241 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     3301 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
     3361 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     3421 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     3481 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
     3541 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     3601 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
     3661 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     3721 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     3781 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     3841 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     3901 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     3961 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     4021 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     4081 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     4141 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     4201 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     4261 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     4321 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     4381 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     4441 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     4501 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     4561 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     4621 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     4681 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     4741 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     4801 gccaagaagg gcggaaagat cgccgtgtaa ttctagatca ctggatttta ggaattagaa
     4861 attttattga tagaagtatt ttacaaatac aaatacatac taagggtttc ttatatgctc
     4921 aacacatgag cgaaacccta taagaaccct aattccctta tctggaattc gatatcaagc
     4981 ttatcgatac cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     5041 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5101 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5161 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgaaattgta
     5221 aacgttaatg ttatcgatac atgagaatta agggagtcac gttatgaccc ccgccgatga
     5281 cgcgggacaa gccgttttac gtttggaact gacagaaccg caacgttgaa ggagccactc
     5341 agccgcgggt ttctggagtt taatgagcta agcacatacg tcagaaacca ttattgcgcg
     5401 ttcaaaagtc gcctaaggtc actatcagct agcaaatatt tcttgtcaaa aatgctccac
     5461 tgacgttcca taaattcccc tcggtatcca attagagtct catattcact ctcaactcga
     5521 tcgaggcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5581 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5641 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5701 tgaactccaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5761 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5821 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5881 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5941 acatcgcatc gagcgaggac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6001 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat
     6061 gcccgacggc gaggatctcg tcgtgaccca gggcgatgcc tgcttgccga atatcatggt
     6121 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6181 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6241 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6301 ccttcttgac gagttcttct gagcgggact ctggggttcg gactctagct agagtcaagc
     6361 agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     6421 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     6481 catgacgtta tttatgagat gggttttttg attagagtcc cgcaattata catttaatac
     6541 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     6601 atgttactag atcgaccggc atgcaagctg ataacgttac accacaatat atcctgccaa
     6661 gatctaattc cggggatcgg aaatccagaa gcccgagagg ttgccgcctt tcgggctttt
     6721 tctttttcaa aaaaaaaaat ttataaaacg atctgttgcg gccggccgcc gggttgtggg
     6781 caaaggcgct ggcgctcgac ggtgggcaac cgcttgcggt tgtccacggg cggagccggt
     6841 gcgcgtagcg cattgtccac aagccaaggg cgaccaataa ttgatatata tattcataat
     6901 tgaaaagcta attgaacata ctacttgctg taactacttg ccggagcgag gggtgtttgc
     6961 aagctgttga tctgaaaggg ctattagcgt tctcacgtgc ctttttgatt agcgatttca
     7021 cgtgacctta ttagcgattt cacgtactcc gattagcgat ttcacgtacc ctgattagcg
     7081 atttcacgtg gatagttttt ggagcgggcc ggaaagcccc gtgaatcaag gctttgcggg
     7141 gcattagcgg tttcacgtgg ataactaccc tctatccaca ggcttccggg gataaaaa
//
