pKeratin-emiRFP2-N1

LOCUS       pKeratin-emiRFP2-N1      6180 bp DNA     circular SYN 01-JUN-2023
DEFINITION  Keratin labeling with near-infrared fluorescent protein 
            miRFP670nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6180)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 6180)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 1, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
FEATURES             Location/Qualifiers
     source          1..6180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             625..1914
                     /codon_start=1
                     /label=Keratin18
                     /translation="MSFTTRSTFSTNYRSLGSVQAPSYGARPVSSAASVYAGAGGSGSR
                     ISVSRSTSFRGGMGSGGLATGIAGGLAGMGGIQNEKETMQSLNDRLASYLDRVRSLETE
                     NRRLESKIREHLEKKGPQVRDWSHYFKIIEDLRAQIFANTVDNARIVLQIDNARLAADD
                     FRVKYETELAMRQSVENDIHGLRKVIDDTNITRLQLETEIEALKEELLFMKKNHEEEVK
                     GLQAQIASSGLTVEVDAPKSQDLAKIMADIRAQYDELARKNREELDKYWSQQIEESTTV
                     LTTQSAEVGAAETTLTELRRTVQSLEIDLDSMRNLKASLENSLREVEARYALQMEQLNG
                     ILLHLESELAQTRAEGQRQAQEYEALLNIKVKLEAEIATYRRLLEDGEDFNLGDALDSS
                     NSMQTIQKTTTRRIVDGKVVSETNDTKVLRH"
     primer_bind     706..722
                     /label=0421-PM398-F
     primer_bind     1785..1812
                     /label=Keratin-F
     primer_bind     1917..1964
                     /label=0421-PM398-F
     CDS             1945..2868
                     /codon_start=1
                     /label=emiRFP2
                     /translation="MAEGSVARQPDLLTCEHEEIHLAGSIQPHGTLLVVSEPDHRIIQA
                     SANAAEFLNLGSVLGIPLAEIDGDLLIKILPHLDLTAEGMPVAVRCRIGNPSTEYDGLM
                     HRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTGYDRVM
                     VYRFDERGHGEVYSEIHVTGLESYFGNRYPSSLVPQMARRLYVRQRVRVLVDVTYQPVP
                     LEPRLSPLTGRDLDMSGCFLRSMSPTHLQFLKNMGVRATLVVSLVVGGKLWGLVICHHY
                     LPRFIHFELRAICVLLAEAIATRITAL"
     regulatory      1945..1948
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     unsure          2069
                     /label=Val
     unsure          2309..2310
                     /label=Ser
     unsure          2392..2394
                     /label=Val
     CDS             2530..2532
                     /label=Thr
     unsure          2560
                     /label=Val
     unsure          2594
                     /label=Thr
     CDS             2686..2688
                     /label=Cys
     unsure          2708
                     /label=Asp
     unsure          2731..2733
                     /label=Ala
     primer_bind     complement(2849..2894)
                     /label=0421-PM398-R
     polyA_signal    2993..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3030..3049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3084..3103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3121..3576)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3258..3279)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3470..3489
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3603..3707
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3709..4066
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3732..3752)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3917..4052
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3979..3998
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(4002..4013)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4101..4895
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4155..4174)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4289..4299
                     /label=0421-PM398-F
     primer_bind     4765..4784
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5083..5102)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5127..5174
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5503..6091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5992..6011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact tttcaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag caccatgagc ttcaccactc gctccacctt ctccaccaac
      661 taccggtccc tgggctctgt ccaggcgccc agctacggcg cccggccggt cagcagcgcg
      721 gccagcgtct atgcaggcgc tgggggctct ggttcccgga tctccgtgtc ccgctccacc
      781 agcttcaggg gcggcatggg gtccgggggc ctggccaccg ggatagccgg gggtctggca
      841 ggaatgggag gcatccagaa cgagaaggag accatgcaaa gcctgaacga ccgcctggcc
      901 tcttacctgg acagagtgag gagcctggag accgagaacc ggaggctgga gagcaaaatc
      961 cgggagcact tggagaagaa gggaccccag gtcagagact ggagccatta cttcaagatc
     1021 atcgaggacc tgagggctca gatcttcgca aatactgtgg acaatgcccg catcgttctg
     1081 cagattgaca atgcccgtct tgctgctgat gactttagag tcaagtatga gacagagctg
     1141 gccatgcgcc agtctgtgga gaacgacatc catgggctcc gcaaggtcat tgatgacacc
     1201 aatatcacac gactgcagct ggagacagag atcgaggctc tcaaggagga gctgctcttc
     1261 atgaagaaga accacgaaga ggaagtaaaa ggcctacaag cccagattgc cagctctggg
     1321 ttgaccgtgg aggtagatgc ccccaaatct caggacctcg ccaagatcat ggcagacatc
     1381 cgggcccaat atgacgagct ggctcggaag aaccgagagg agctagacaa gtactggtct
     1441 cagcagattg aggagagcac cacagtgctc accacacagt ctgctgaggt tggagctgct
     1501 gagacgacgc tcacagagct gagacgtaca gtccagtcct tggagatcga cctggactcc
     1561 atgagaaatc tgaaggccag cttggagaac agcctgaggg aggtggaggc ccgctacgcc
     1621 ctacagatgg agcagctcaa cgggatcctg ctgcaccttg agtcagagct ggcacagacc
     1681 cgggcagagg gacagcgcca ggcccaggag tatgaggccc tgctgaacat caaggtcaag
     1741 ctggaggctg agatcgccac ctaccgccgc ctgctggaag atggcgagga ctttaatctt
     1801 ggtgatgcct tggacagcag caactccatg caaaccatcc aaaagaccac cacccgccgg
     1861 atagtggatg gcaaagtggt gtctgagacc aatgacacca aagttctgag gcatcgaatt
     1921 ctgcagtcga cggtaccacc ggtcatggcc gagggcagcg tggccagaca gcccgacctg
     1981 ctgacctgtg agcacgaaga aatccatctg gccggatcca tccagccaca cggaacactg
     2041 ctggtggtct ccgagcctga ccatagaata attcaggcat ctgccaacgc cgctgaattc
     2101 ctgaatctgg gaagtgtgct gggaatcccc ctggctgaga tcgacgggga tctactgatc
     2161 aagattctgc cccacctgga ccttaccgca gaaggaatgc cagtggccgt caggtgccgc
     2221 attggaaacc cctccacaga gtacgacgga ctgatgcatc gaccacctga aggaggactg
     2281 atcattgagc tggaacgggc tgggccatcc attgatctga gcggaacact ggctcccgca
     2341 ctggagcgaa ttcgaactgc aggatccctg agggctctgt gcgacgatac tgtactgctg
     2401 ttccagcagt gtaccggcta cgaccgagtg atggtctatc ggttcgatga acgggggcac
     2461 ggagaggtgt attcagaaat tcatgtgact ggactggaga gctactttgg caacaggtat
     2521 cctagctccc tcgtgccaca gatggcccgg cggctgtacg tgagacagag ggtgcgcgtc
     2581 ctggtagatg tcacctatca gcctgtgcca ctggagccaa gactgagtcc cctgactggc
     2641 agggacctgg atatgtcagg gtgttttctg cgctctatga gtcctactca cctgcagttc
     2701 ctgaagaaca tgggcgtgcg ggcaacactg gtggtcagcc tggtggtcgg aggcaaactg
     2761 tgggggctgg tgatctgcca ccattatctg ccacgcttca tccattttga gctgcgagca
     2821 atttgtgtgc tgctggccga agctatcgca acccgcatta cagccctgta ggcggccgcg
     2881 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2941 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3121 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3181 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3241 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3301 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3361 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3421 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3481 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3541 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3601 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3661 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3721 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3781 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3841 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3901 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3961 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4021 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4081 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4141 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4201 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4261 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4321 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4381 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4441 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4501 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4561 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4621 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4681 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4741 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4801 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4861 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4921 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4981 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5041 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5101 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5161 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5221 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5281 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5341 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5401 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5461 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5521 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5581 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5641 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5701 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5761 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5821 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5881 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5941 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6001 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6061 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6121 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
//