LOCUS       sequence-356873-        5457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the reversibly switchable fluorescent protein Padron2 in 
            bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  AUTHORS   Konen T, Stumpf D, Grotjohann T, Jansen I, Bossi M, Weber M, Jensen 
            N, Hell SW, Jakobs S
  TITLE     The Positive Switching Fluorescent Protein Padron2 Enables Live-Cell
            Reversible Saturable Optical Linear Fluorescence Transitions 
            (RESOLFT) Nanoscopy without Sequential Illumination Steps.
  JOURNAL   ACS Nano. 2021 May 21. doi: 10.1021/acsnano.0c08207.
  PUBMED    34019380
REFERENCE   2  (bases 1 to 5457)
  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: "ACS Nano. 
            2021 May 21. doi: 10.1021/acsnano.0c08207."
COMMENT     Sequence Label: pBAD-Padron2
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             312..334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             354..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             375..407
                     /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             420..440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             474..1142
                     /codon_start=1
                     /product="fast-switching photoswitchable fluorescent 
                     protein useful for fluorescence nanoscopy (Tiwari et al., 
                     2015)"
                     /label=Kohinoor
                     /translation="MAVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSVDLKVK
                     EGGPLPFAYDILSMAFCYGNKVFIKYPENIVDYFKQLFPEGYSWERSMIYEDGGICNAT
                     NDITLDGDCFIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKSDGNYALSL
                     EGGGHYRCDSKTTYKAKKVVQLPDYHAVDHHIEIKSHDKDYSNVNLHEHAEAHSGLPGQ
                     A"
     primer_bind     complement(1240..1257)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1410..1496
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1588..1615
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1634..1725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1726..1794,1795..2586)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGEPLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1944..1963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2628..3083
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2715..2734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2925..2946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3194..3782
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3683..3702
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3936..3953
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3968..4108
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4034..4053)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4194..4216
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4354..4373
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4553..5431)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPPRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atactacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc
      361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
      421 agaacctgta cttccagggc tcgagcatgg tgagcaaggg cgaggagaac aacatggccg
      481 tgattaaacc agacatgaag atcaagctgc gtatggaagg cgctgtaaat ggacacccgt
      541 tcgcgattga aggagttggc cttgggaagc ctttcgaggg aaaacagagt gtggacctta
      601 aagtcaaaga aggcggacct ctgcctttcg cctatgacat cttgtccatg gcgttctgtt
      661 acggcaacaa ggtattcatc aaatacccag aaaatatagt agactatttc aagcagcttt
      721 ttcctgaggg ctactcttgg gaacgaagca tgatttacga agacgggggc atttgtaacg
      781 cgacaaacga cataaccctg gatggtgact gttttatcta tgaaattcga tttgatggtg
      841 tgaactttcc tgccaatggt ccagttatgc agaagaggac tgtgaaatgg gagccatcca
      901 ctgagaaatt gtatgtgcgt gatggagtgc tgaagtctga tggtaactac gctctgtcgc
      961 ttgaaggagg tggccattac cgatgtgact ccaaaactac ttataaagct aagaaggttg
     1021 tccagttgcc agactatcac gctgtggacc accacattga gattaaaagc cacgacaaag
     1081 attacagtaa tgttaatctg catgagcatg ccgaagcgca ttctgggctg ccggggcagg
     1141 ccatggacga gctgtacaag taagaattcg aagatggacg agctgtacaa gtaagaattc
     1201 gaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     1261 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     1321 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     1381 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1441 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1501 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggta ggacgcccgc
     1561 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     1621 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     1981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     2281 acaacgttgc gcaaactatt aactggcgaa ccacttactc tagcttcccg gcaacaatta
     2341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag
     2641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2701 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2761 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2821 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2941 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     3001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3061 caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3121 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3181 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3241 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3301 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3361 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3421 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3481 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3541 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3601 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3661 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3721 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3781 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3841 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3901 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3961 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatct
     4021 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     4081 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     4141 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4201 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcaga aggagatggc
     4261 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     4321 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     4381 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
     4441 atgtttgaca gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc
     4501 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
     4561 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
     4621 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
     4681 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
     4741 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
     4801 gacgcggcgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
     4861 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
     4921 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
     4981 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
     5041 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
     5101 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
     5161 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
     5221 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
     5281 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
     5341 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
     5401 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagag
//
