LOCUS       sequence-121993-        7130 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 7130)
  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 7130)
  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: pOPTXLUC
FEATURES             Location/Qualifiers
     source          1..7130
                     /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(3081..3105)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3110..4762
                     /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(3176..3194)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4917..4933)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4918..4934)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4958..4977)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4959..4977)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4984..5001)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4984..5000)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4993..5015)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        5247..5426
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5460..6254
                     /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(5514..5533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6124..6143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6294..6545
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    6569..6591
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      join(6724..7130,1..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 aaattaatgt agaaatagtt gattaatatt ggataatatt tttctagttt ctctttcatg
     2941 attcttttaa aaatgaacga gacattataa gtacctcctc gctccccacc tagttaattt
     3001 aactttgtgc caactcctaa agccaaactt atatcttcct ttgtttgaca taatacaaga
     3061 aaaaaagaaa agagctatgg acccagcttt cttgtacaaa gtggttgcca tggaagacgc
     3121 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
     3181 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
     3241 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
     3301 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     3361 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     3421 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     3481 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     3541 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     3601 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     3661 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     3721 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     3781 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     3841 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     3901 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     3961 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     4021 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     4081 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     4141 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     4201 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     4261 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     4321 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     4381 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     4441 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     4501 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     4561 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     4621 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     4681 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     4741 gggcggaaag atcgccgtgt aattctagat cactggattt taggaattag aaattttatt
     4801 gatagaagta ttttacaaat acaaatacat actaagggtt tcttatatgc tcaacacatg
     4861 agcgaaaccc tataagaacc ctaattccct tatctggaat tcgatatcaa gcttatcgat
     4921 accgtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacaa
     4981 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     5041 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     5101 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcgaaattg taaacgttaa
     5161 tgttatcgat acatgagaat taagggagtc acgttatgac ccccgccgat gacgcgggac
     5221 aagccgtttt acgtttggaa ctgacagaac cgcaacgttg aaggagccac tcagccgcgg
     5281 gtttctggag tttaatgagc taagcacata cgtcagaaac cattattgcg cgttcaaaag
     5341 tcgcctaagg tcactatcag ctagcaaata tttcttgtca aaaatgctcc actgacgttc
     5401 cataaattcc cctcggtatc caattagagt ctcatattca ctctcaactc gatcgaggca
     5461 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5521 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5581 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactcc
     5641 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5701 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5761 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5821 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5881 tcgagcgagg acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5941 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgg atgcccgacg
     6001 gcgaggatct cgtcgtgacc cagggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6061 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6121 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6181 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6241 acgagttctt ctgagcggga ctctggggtt cggactctag ctagagtcaa gcagatcgtt
     6301 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta
     6361 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
     6421 tatttatgag atgggttttt tgattagagt cccgcaatta tacatttaat acgcgataga
     6481 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     6541 agatcgaccg gcatgcaagc tgataacgtt acaccacaat atatcctgcc aagatctaat
     6601 tccggggatc ggaaatccag aagcccgaga ggttgccgcc tttcgggctt tttctttttc
     6661 aaaaaaaaaa atttataaaa cgatctgttg cggccggccg ccgggttgtg ggcaaaggcg
     6721 ctggcgctcg acggtgggca accgcttgcg gttgtccacg ggcggagccg gtgcgcgtag
     6781 cgcattgtcc acaagccaag ggcgaccaat aattgatata tatattcata attgaaaagc
     6841 taattgaaca tactacttgc tgtaactact tgccggagcg aggggtgttt gcaagctgtt
     6901 gatctgaaag ggctattagc gttctcacgt gcctttttga ttagcgattt cacgtgacct
     6961 tattagcgat ttcacgtact ccgattagcg atttcacgta ccctgattag cgatttcacg
     7021 tggatagttt ttggagcggg ccggaaagcc ccgtgaatca aggctttgcg gggcattagc
     7081 ggtttcacgt ggataactac cctctatcca caggcttccg gggataaaaa
//
