LOCUS       sequence-378486-        4784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmids for pH sensitive fluorescent protein 
            mLemon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4784)
  TITLE     Bacterial expression plasmids for pH sensitive fluorescent proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4784)
  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            "Unpublished"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-mLemon
FEATURES             Location/Qualifiers
     source          1..4784
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      32..59
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        78..169
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(170..238,239..1030)
                     /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(388..407)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1201..1789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1690..1709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1943..1960
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1975..2115
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2041..2060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2201..2223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2379..3257)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3284..3568
                     /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     3491..3510
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3614..3631
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3635..3667
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(3671..3679,3680..3694)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(4468..4485)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4638..4724
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
       61 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      121 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      181 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      241 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      301 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      361 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
      421 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      481 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      541 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      601 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      661 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
      721 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
      781 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
      841 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
      901 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
      961 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1021 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1081 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1141 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1201 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaacaac caccgctacc
     1261 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1321 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     1381 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1441 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1501 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1561 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1621 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1681 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1741 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1801 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1861 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     1921 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     1981 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     2041 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     2101 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2161 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2221 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg
     2281 aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc
     2341 tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca
     2401 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     2461 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     2521 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     2581 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     2641 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     2701 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     2761 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     2821 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     2881 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     2941 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     3001 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     3061 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     3121 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     3181 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     3241 ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca
     3301 tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg
     3361 gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg
     3421 cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc
     3481 acactttgct atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt
     3541 ttatcgcaac tctctactgt ttctccatac ccgttttttg ggctaacagg aggaattaac
     3601 catggggggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
     3661 aatgggtcgg gatctgtacg acgatgacga taaggatccg agctcgagaa tggtgagtaa
     3721 aggagaagaa cttttcactg gagtggtgcc tatcctggtg gaactggatg gcgacgtgaa
     3781 cggccacaag ttcagcgtta gaggcgaagg cgaaggggat gccacaaacg gcaagctgac
     3841 cctgaagtta atcagcacca caggcaagct gcccgtgcct tggcctacac tggtcacaac
     3901 actgggctac ggcgtgcaga gcttcgccag ataccccgac cacatgaagc ggcacgattt
     3961 cttcaagagc gccatgccag agggctacgt gcaagagaga accatcagct tcaaggacga
     4021 cggcacctac aagaccagag ccgaagtgaa gttcgagggc gacaccctgg tcaaccggat
     4081 cgagctgaag ggcatcgact tcaaagagga cggcaacatc ctgggccaca aacttgagta
     4141 caacttcaac gaccaccagg tctacatcac cgccgacaag cagaagaacg gcatcaaggc
     4201 caacttcaag atccggcaca acgtggaaga tggcagcgtg cagctggccg atcactacca
     4261 gcagaacaca cccatcggag atggccctgt gctgctgccc gataaccact acctgttcta
     4321 taccagcaca ctgagcaagg accccaacga gaagcgggac cacatggtgc tgaaggaatt
     4381 tgtgaccgcc gccggcatta cacatggcat ggatgaacta tacaaatgaa agcttggctg
     4441 ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg
     4501 tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc
     4561 gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt
     4621 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     4681 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     4741 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgc
//
