LOCUS       sequence-178732-        5732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP in the presence of ipgC and mxiE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  TITLE     Unpublished Murray Lab Plasmids
REFERENCE   2  (bases 1 to 5732)
  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"
COMMENT     Sequence Label: PipaH-deGFP
FEATURES             Location/Qualifiers
     source          1..5732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             366..388
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(425..446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(686..705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1033..1054
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1149..1169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1574..2368)
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1685..1704)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2295..2314
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      3848..4617
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             4618..5280
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 gatccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
       61 aggctagggt caaaaatcgt ggcgttgaca aaatggctgc gttacgtcat tgagcatatc
      121 caggactggc cggcaaaccg ggtacgcgat ctgttgcctt ggaaagttga tctgacctct
      181 cagtaaatat caatacggtt ctgacgagcc gcttaccgtt caaatatgaa gtacgatgtt
      241 taactaaccg aaaaacaaga acaatacggt gcaaacaggc cattcacggt taactgaaac
      301 agtatcgttt ttttacagcc aattttgttt atccttatta agataaaaaa gtgctaagca
      361 ataattttgt ttaactttaa gaaggagata tacaatggag cttttcactg gcgttgttcc
      421 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      481 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      541 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      601 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      661 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      721 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      781 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      841 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
      901 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      961 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1021 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatct aatgaagcat
     1081 ctggtgaata actcgagcaa agcccgccga aaggcgggct tttctgtgtc gaccgatgcc
     1141 cttactagtg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1201 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1261 tgcgtattgg gcgcatgcat aaaaactgtt gtaattcatt aagcattctg ccgacatgga
     1321 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     1381 gcgtataata tttgcccatg ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
     1441 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
     1501 gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac
     1561 cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     1621 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     1681 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     1741 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     1801 cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca
     1861 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     1921 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     1981 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     2041 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     2101 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     2161 gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac aggtcggtct
     2221 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     2281 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     2341 ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag
     2401 attttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     2461 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc
     2521 ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct
     2581 ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc ccaggtggca
     2641 cttttcgggg aaatgtgcgc gcccgcgttc ctgctggcgc tgggcctgtt tctggcgctg
     2701 gacttcccgc tgttccgtca gcagcttttc gcccacggcc ttgatgatcg cggcggcctt
     2761 ggcctgcata tcccgattca acggccccag ggcgtccaga acgggcttca ggcgctcccg
     2821 aaggtctcgg gccgtctctt gggcttgatc ggccttcttg cgcatctcac gcgctcctgc
     2881 ggcggcctgt agggcaggct catacccctg ccgaaccgct tttgtcagcc ggtcggccac
     2941 ggcttccggc gtctcaacgc gctttgagat tcccagcttt tcggccaatc cctgcggtgc
     3001 ataggcgcgt ggctcgaccg cttgcgggct gatggtgacg tggcccactg gtggccgctc
     3061 cagggcctcg tagaacgcct gaatgcgcgt gtgacgtgcc ttgctgccct cgatgccccg
     3121 ttgcagccct agatcggcca cagcggccgc aaacgtggtc tggtcgcggg tcatctgcgc
     3181 tttgttgccg atgaactcct tggccgacag cctgccgtcc tgcgtcagcg gcaccacgaa
     3241 cgcggtcatg tgcgggctgg tttcgtcacg gtggatgctg gccgtcacga tgcgatccgc
     3301 cccgtacttg tccgccagcc acttgtgcgc cttctcgaag aacgccgcct gctgttcttg
     3361 gctggccgac ttccaccatt ccgggctggc cgtcatgacg tactcgaccg ccaacacagc
     3421 gtccttgcgc cgcttctctg gcagcaactc gcgcagtcgg cccatcgctt catcggtgct
     3481 gctggccgcc cagtgctcgt tctctggcgt cctgctggcg tcagcgttgg gcgtctcgcg
     3541 ctcgcggtag gcgtgcttga gactggccgc cacgttgccc attttcgcca gcttcttgca
     3601 tcgcatgatc gcgtatgccg ccatgcctgc ccctcccttt tggtgtccaa ccggctcgac
     3661 gggggcagcg caaggcggtg cctccggcgg gccactcaat gcttgagtat actcactaga
     3721 ctttgcttcg caaagtcgtg accgcctacg gcggctgcgg cgccctacgg gcttgctctc
     3781 cgggcttcgc cctgcgcggt cgctgcgctc ccttgccagc ccgtggatat gtggacgatg
     3841 gccgcgagcg gccaccggct ggctcgcttc gctcggcccg tggacaaccc tgctggacaa
     3901 gctgatggac aggctgcgcc tgcccacgag cttgaccaca gggattgccc accggctacc
     3961 cagccttcga ccacataccc accggctcca actgcgcggc ctgcggcctt gccccatcaa
     4021 tttttttaat tttctctggg gaaaagcctc cggcctgcgg cctgcgcgct tcgcttgccg
     4081 gttggacacc aagtggaagg cgggtcaagg ctcgcgcagc gaccgcgcag cggcttggcc
     4141 ttgacgcgcc tggaacgacc caagcctatg cgagtggggg cagtcgaagg cgaagcccgc
     4201 ccgcctgccc cccgagcctc acggcggcga gtgcgggggt tccaaggggg cagcgccacc
     4261 ttgggcaagg ccgaaggccg cgcagtcgat caacaagccc cggaggggcc actttttgcc
     4321 ggagggggag ccgcgccgaa ggcgtggggg aaccccgcag gggtgccctt ctttgggcac
     4381 caaagaacta gatatagggc gaaatgcgaa agacttaaaa atcaacaact taaaaaaggg
     4441 gggtacgcaa cagctcattg cggcaccccc cgcaatagct cattgcgtag gttaaagaaa
     4501 atctgtaatt gactgccact tttacgcaac gcataattgt tgtcgcgctg ccgaaaagtt
     4561 gcagctgatt gcgcatggtg ccgcaaccgt gcggcaccct accgcatgga gataagcatg
     4621 gccacgcagt ccagagaaat cggcattcaa gccaagaaca agcccggtca ctgggtgcaa
     4681 acggaacgca aagcgcatga ggcgtgggcc gggcttattg cgaggaaacc cacggcggca
     4741 atgctgctgc atcacctcgt ggcgcagatg ggccaccaga acgccgtggt ggtcagccag
     4801 aagacacttt ccaagctcat cggacgttct ttgcggacgg tccaatacgc agtcaaggac
     4861 ttggtggccg agcgctggat ctccgtcgtg aagctcaacg gccccggcac cgtgtcggcc
     4921 tacgtggtca atgaccgcgt ggcgtggggc cagccccgcg accagttgcg cctgtcggtg
     4981 ttcagtgccg ccgtggtggt tgatcacgac gaccaggacg aatcgctgtt ggggcatggc
     5041 gacctgcgcc gcatcccgac cctgtatccg ggcgagcagc aactaccgac cggccccggc
     5101 gaggagccgc ccagccagcc cggcattccg ggcatggaac cagacctgcc agccttgacc
     5161 gaaacggagg aatgggaacg gcgcgggcag cagcgcctgc cgatgcccga tgagccgtgt
     5221 tttctggacg atggcgagcc gttggagccg ccgacacggg tcacgctgcc gcgccggtag
     5281 cacttgggtt gcgcagcaac ccgtaagtgc gctgttccag actatcggct gtagccgcct
     5341 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     5401 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccgtttt tatcaggctc
     5461 tgggaggcag aataaatgat catatcgtca attattacct ccacggggag agcctgagca
     5521 aactggcctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     5581 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     5641 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     5701 accaggcgtt taagggcacc aataactgcc tg
//
