LOCUS       sequence-352983-        7264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7264)
  AUTHORS   Kanther M, Sun X, Muhlbauer M, Mackey LC, Flynn EJ 3rd, Bagnat M, 
            Jobin C, Rawls JF
  TITLE     Microbial colonization induces dynamic temporal and spatial patterns
            of NF-kappaB activation in the zebrafish digestive tract.
  JOURNAL   Gastroenterology. 2011 Jul;141(1):197-207. doi: 
            10.1053/j.gastro.2011.03.042. Epub 2011 Mar 24.
  PUBMED    21439961
REFERENCE   2  (bases 1 to 7264)
  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: 
            "10.1053/j.gastro.2011.03.042"; journalName: "Gastroenterology"; 
            date: "2011-07"; volume: "141"; issue: "1"; pages: "197-207"
COMMENT     Sequence Label: pNFkB:eGFP
FEATURES             Location/Qualifiers
     source          1..7264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    116..137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(152..157,158..175,176..182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    190..206
                     /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     195..217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     214..230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     214..230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     249..269
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        251..269
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     306..322
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     306..322
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(356..367,368..555)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     polyA_signal    994..1128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1031..1050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1085..1104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1173..1886,1887..1889,1890..1892))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1176..1886,1887..1889,1890..1892))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1218..1239)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1567..1586
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1826..1847
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1889..1898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1959..1970
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    2136..2160
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2298..2322)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(2701..2863,2864..2875)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4700..4719)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4701..4719)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4729..4746)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4729..4745)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4738..4760)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4887..5342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4974..4993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5184..5205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5368..5472
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5473..5541,5542..6333)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5691..5710)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6504..7092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6993..7012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7246..7263
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
       61 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      121 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      181 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      241 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg
      301 gccgctctag aactagtgga tctgctgggc ttgctgaagg tagggggtca agaaccagag
      361 gtgtaaagta cttgagtaat tttacttgat tactgtactt aagtattatt tttggggatt
      421 tttactttac ttgagtacaa ttaaaaatca atacttttac ttttacttaa ttacattttt
      481 ttagaaaaaa aagtactttt tactccttac aattttattt acagtcaaaa agtacttatt
      541 ttttggagat cacttcattc tattttccct tgctattacc aaaccaattg aattgcgctg
      601 atgcccagtt taatttaaat gttatttatt ctgcctatga aaatcgtttt cacattatat
      661 gaaattggtc agacatgttc attggtcctt tggaagtgac gtcatgtcac atctattacc
      721 acaatgcaca gcaccttgac ctggaaatta gggaaattat aacagtcaat cagtggaaga
      781 aaatggagga agtatgtgat tcatcagcag ctgcgagcag cacagtccaa aatcagccac
      841 aggatcaaga gcacccgtgg ccgtatcttc gcagatctga tctagaggat cataatcagc
      901 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
      961 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1021 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1081 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcat catcgatgaa
     1141 ttaattcgag ctccaccgcg gtggcggccg ctttacttgt acagctcgtc catgccgaga
     1201 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     1261 tctttgctca gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     1321 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
     1381 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     1441 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     1501 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     1561 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     1621 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     1681 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     1741 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     1801 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     1861 ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtggatc ctcgaagttt
     1921 ggggaaagcc cggcaagggg tccaggggta gacactggtg ggagctgcag agcagagctg
     1981 ggtgggagcg cggtcactgc tcgttcgcgg aaccgccggc tctatccagt cttctcagtt
     2041 gctcgctgca gtcgcggttg gagtagtagg cgcctcagct ggcgccttta tagaagcgct
     2101 gtgaatggat ggacttccta cgtcactgga tatcaaccac tttgtacaag aaagctgggt
     2161 cgagctcaag cttcgaattc tgcaggtcga cggaaagtcc ccagcggaaa gtccctcgac
     2221 ggaaagtccc cagcggaaag tccctcgacg gaaagtcccc agcggaaagt ccctcgaccg
     2281 tgcatgcatt cacaggaagc ctgctttttt gtacaaactt gttgatatct cgagggccca
     2341 tctggcctgt gtttcagaca ccagggagtc tctgctcacg tttcctgcta tttgcagcct
     2401 ctctatcaag actaatacac ctcttcccgc atcggctgcc tgtgagaggc ttttcagcac
     2461 tgcaggattg cttttcagcc ccaaaagagc taggcttgac actaacaatt ttgagaatca
     2521 gcttctactg aagttaaatc tgaggtttta caactttgag tagcgtgtac tggcattaga
     2581 ttgtctgtct tatagtttga taattaaata caaacagttc taaagcagga taaaaccttg
     2641 tatgcatttc atttaatgtt ttttgagatt aaaagcttaa acaagaatct ctagttttct
     2701 ttcttgcttt tacttttact tccttaatac tcaagtacaa ttttaatgga gtactttttt
     2761 acttttactc aagtaagatt ctagccagat acttttactt ttaattgagt aaaattttcc
     2821 ctaagtactt gtactttcac ttgagtaaaa tttttgagta ctttttacac ctctgtcaag
     2881 aactcctgga caaacctctg acctgtgtgg aacagagtgg atatgggtgt ctgaacagat
     2941 attcacgtct tttgcagatc agagggcatt tctggtgttg tgcaatgacc tgggtccaac
     3001 aggtaggcgt actcgtatcc aagagcattg ttggacagga ggtagtctcc attcctataa
     3061 agaggaagga atggcaccat gtagtagccg tcattgtggc caattggtgc attggtacgt
     3121 gggtagatgg accggggagg ctgatgagtt cttagccagc gctcaaagat gctagaaatg
     3181 atgtaagaaa caaatatgca agatgaaagt taagataaga taaaaaatta aacaaataaa
     3241 tgcaattatg ttttgtgttt ctattccacc aataataacc tcacgcagta aagccaccag
     3301 attatttttt gttctttgct ctgcagggca accaaactct aacaattgat ctcaattgat
     3361 ctttcgctgc tcatccaagt acattcacaa acttcactaa ttgtagcagc tataaacaac
     3421 agattgtatg aactttgtca aaaactgtgc agtctaatgt catgacgtgt ttgttacctg
     3481 tcaatgaaag catggtgaag gaggaagatg gggtcgttgg ctgagccctg gactgaggac
     3541 attgagccgt tcatgaagac atgcagcgca ttgtgcatgg tgctctggcc ttgcacagcc
     3601 atgcctgttt ctgggctggc aaaacctaca aagaaaagaa gcaggtgaag aagttacagt
     3661 ttcaatcaag ataacagcaa aacaaggcta attaaaagac agcaaaaaaa ggagatgtaa
     3721 aatatagcta ttgattttct ttacttttct ttacgtaggc aaatgtgact taagtgtgtt
     3781 gttgtatccc cccccccccc tttctgtaat gaatctgtgg aaattatttc ttattttgac
     3841 taccaatcaa tcaatcaatc aatcaatcaa tcaatcaatc aatcaatcaa tcaatcaatc
     3901 aatcaatcaa tcaatcaatc taccaaccaa ccaaccaacc aacgactgta tatgagaact
     3961 gtaatgagtg acctcctcct tctagtgatc caaacaggaa gaagctccaa atggcatgaa
     4021 tgctccagtg gtggcattca tattaaaatc tttatttaac atgtacttgt taatcataat
     4081 ttctgtaata acttttcttt tctgtagccc aagttgttga tattataaac tgaccaacca
     4141 gatgccttaa taaaagtatg tgctctcaca tccaacgttc aaaacgtttg acagattttg
     4201 tgcagcccgc tttcaagtgg taggctgtgg acttccaaca agcttacttc tgattggtta
     4261 gaggggttgc cataggaacg gtgactcaga ctcgacgcat ctcggctctt tcttgcttat
     4321 tggcaacgtc tagctccaac acggcgatgt ccatacagtg aaaaatggca actgaattaa
     4381 cttcattttg ttggagccag aagtaagtca ttttgaatgg gtgacgtaac actcactcgg
     4441 tccagttctc aaatacagtc aacaagtact attacataat aatagtgtgt actactatta
     4501 catattattg taataatgtc tgttaaacac agtcataatt gaaaaaaaac aactaaaaca
     4561 tttattcaag ggtaaagaca aatgcagaca aatacagaaa agatttaaaa tagcagtaga
     4621 aaatggcaaa cgggcttatt catccaagtt ttcagcactt gtacaggtgg gtaattccta
     4681 ctatttggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     4741 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     4801 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     4861 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     4921 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4981 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5041 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5101 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5161 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5221 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5281 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5341 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5401 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5461 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5521 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6301 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7141 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7201 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7261 gagg
//
