LOCUS       sequence-378487-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmids for pH sensitive fluorescent protein 
            pHmGL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  TITLE     Bacterial expression plasmids for pH sensitive fluorescent proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4719)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBAD-pHmGL
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(172..1050)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1077..1361
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1284..1303
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1404..1421
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1431..1433,1434..1436,1437..2150)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFVDHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(1476..1497)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2196..2213)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2366..2452
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2544..2571
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2590..2681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2682..2750,2751..3542)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2900..2919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3713..4301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4202..4221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4455..4472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4487..4627
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4553..4572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(4713..4719,1..16)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc
       61 gcgcgcgaag gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct
      121 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
      181 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
      241 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
      301 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
      361 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
      421 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
      481 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
      541 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
      601 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
      661 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
      721 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
      781 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
      841 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
      901 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
      961 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     1021 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     1081 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     1141 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     1201 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     1261 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     1321 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt ttgggctaac
     1381 aggaggaatt aaccatgggg agccaccatc accatcacca tggcagatct atggtgagca
     1441 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1501 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     1561 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1621 ccttaggcta cggcgtggcc tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1681 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     1741 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1801 tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1861 acaacttcgt tgaccataag gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     1921 ctaacttcaa gacccgccac aacgttgagg acggcggcgt gcagctcgcc gaccactacc
     1981 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     2041 atcagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgaaggaga
     2101 gggtgaccgc cgccgggatt acacatgaca tggacgagct gtacaagtaa gaattcgaag
     2161 cttggctgtt ttggtggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     2221 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     2281 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     2341 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2401 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2461 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2521 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     2581 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2641 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2701 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2761 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2821 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2881 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2941 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3001 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3061 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3121 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3181 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3241 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3301 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3361 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3421 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3481 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3541 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3601 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3661 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3721 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3781 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3841 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3901 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3961 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4021 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4081 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4141 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4201 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4261 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4321 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4381 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4441 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4501 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     4561 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4621 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4681 ggcatccgct tacagacaag ctgtgaccgt ctccgggag
//
