LOCUS       sequence-484240-        5417 bp    DNA     circular SYN 20-AUG-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 588, Emission:
            635.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5417)
  AUTHORS   Shcherbo D, Merzlyak EM, Chepurnykh TV, Fradkov AF, Ermakova GV, 
            Solovieva EA, Lukyanov KA, Bogdanova EA, Zaraisky AG, Lukyanov S, 
            Chudakov DM
  TITLE     Bright far-red fluorescent protein for whole-body imaging.
  JOURNAL   Nat Methods. 2007 Sep . 4(9):741-6.
  PUBMED    17721542
REFERENCE   2  (bases 1 to 5417)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2007 Sep . 4(9):741-6."
COMMENT     Sequence Label: dKatushka-pBAD
FEATURES             Location/Qualifiers
     source          1..5417
                     /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             447..1154
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     primer_bind     complement(1200..1217)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1370..1456
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1548..1575
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1594..1685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1686..1754,1755..2546)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1904..1923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2588..3043
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2675..2694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2885..2906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3643..3662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3896..3913
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3928..4068
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3994..4013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4154..4176
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4314..4333
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4513..5391)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     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 tgggtgagga tagcgtgctg atcaccgaga
      481 acatgcacat gaaactgtac atggagggca ccgtgaacga ccaccacttc aagtgcacat
      541 ccgagggcga aggcaagccc tacgagggca cccagaccat gaagatcaag gtggtcgagg
      601 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa
      661 cctttatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
      721 tcacatggga gaggatcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
      781 ccagcctcca gaacggctgc ctcatctaca acgtcaagat caacggggtg aacttcccat
      841 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccagcacc gagatgctgt
      901 accccgctga cagcggcctg agaggccata gccagatggc cctgaagctc gtgggcgggg
      961 gctacctgca ctgctccctc aagaccacat acagatccaa gaaacccgct aagaacctca
     1021 agatgcccgg cttctacttc gtggacagga gactggaaag aatcaaggag gccgacaaag
     1081 agacctacgt cgagcagcac gagatggctg tggccaggta ctgcgacctg cctagcaaac
     1141 tggggcacag ctgagaattc gaagcttggc tgttttggcg gatgagagaa gattttcagc
     1201 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     1261 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     1321 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     1381 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     1441 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     1501 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     1561 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     1621 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1681 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1741 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1801 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1861 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1921 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1981 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2041 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2101 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2161 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2221 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2281 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2341 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2401 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2461 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2521 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2581 ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2761 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     2821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     2941 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3001 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa
     3061 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3841 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3901 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     3961 cggtatttca caccgcatct ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4021 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4081 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4141 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4201 gcgaggcaga aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     4261 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     4321 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     4381 ccggcgtaga ggatctgctc atgtttgaca gcttatcatc gatgcataat gtgcctgtca
     4441 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     4501 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4561 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4621 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4681 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4741 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     4801 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     4861 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     4921 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     4981 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     5041 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     5101 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     5161 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     5221 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     5281 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     5341 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     5401 actcccgcca ttcagag
//
