LOCUS       sequence-368229-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LSSA12 fluorescent protein (ex/em = 398/511 nm): bacterial 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
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: "Nat 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: pBAD-LSSA12
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..332
                     /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     255..274
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             375..392
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(447..468)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1167..1184)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1337..1423
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1515..1542
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1561..1652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1653..1721,1722..2513)
                     /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(1871..1890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2684..3272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3173..3192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3426..3443
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3458..3598
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3524..3543)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3684..3706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(3862..4719,1..21))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg
       61 tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa
      121 accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa
      181 aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg
      241 cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg
      301 ctttttatcg caactctcta ctgtttctcc atacccgttt tttgggctaa caggaggaat
      361 taaccatggg gagccaccat caccatcacc atggcagatc tatggtgagc aagggcgagg
      421 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      481 agttcagcgt ccgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagc
      541 tcatctccac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttaagct
      601 acggcctgat ggtgttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      661 ccgccatgcc cgaaggctac gtccaggagc gcaccatctc tttcgaggac gacggttact
      721 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgtgctga
      781 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca
      841 acagccacaa cgtctatatc acggccgaca agcagaagaa cggcatcaag gctaacttca
      901 agatccgcca caacgttgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
      961 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc atcgaaagcg
     1021 tactgagcaa agaccccaac gagaagcgcg atcacatggt cctgaaggac ttcgtgaccg
     1081 ccgccgggat tacacatgac atgaacgagc tgtacaagta agaattcgaa gcttggctgt
     1141 tttggtggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1201 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1261 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1321 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1381 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1441 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1501 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1561 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1621 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1681 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1741 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1801 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1861 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     1921 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1981 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2041 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2101 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2161 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2221 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2281 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2341 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2401 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2461 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2521 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2581 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2641 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2701 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2761 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2821 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2881 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2941 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3001 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3061 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3301 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3361 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3421 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3481 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3541 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3601 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3661 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3721 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     3781 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     3841 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     3901 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     3961 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4021 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4081 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4141 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4201 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4261 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4321 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     4381 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     4441 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     4501 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     4561 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     4621 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     4681 caccagatgg gcattaaacg agtatcccgg cagcagggg
//
