LOCUS       sequence-389473-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of Keratin-miRFP670-2 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6189)
  AUTHORS   Zhang H, Papadaki S, Sun X, Wang X, Drobizhev M, Yao L, Rehbock M, 
            Koster RW, Wu L, Namikawa K, Piatkevich KD
  TITLE     Quantitative assessment of near-infrared fluorescent proteins.
  JOURNAL   Nat Methods. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z.
  PUBMED    37666982
REFERENCE   2  (bases 1 to 6189)
  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. 2023 Sep 4. doi: 10.1038/s41592-023-01975-z."
COMMENT     Sequence Label: pKeratin-miRFP670-2-N1
FEATURES             Location/Qualifiers
     source          1..6189
                     /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             1945..2880
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAKRFAERLSTHFTAAHHQR"
     polyA_signal    3002..3123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3039..3058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3093..3112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3130..3585)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3267..3288)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3479..3498
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3612..3716
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3718..4075
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3718..3909
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3741..3761)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3926..4061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4110..4904
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4110..4904
                     /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(4164..4183)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4774..4793
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5092..5111)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5136..5183
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5512..6100
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6001..6020
                     /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 ggtcatggcg cgtaaggtcg atctcacctc ctgcgatcgc
     1981 gagccgatcc acatccccgg cagcattcag ccgtgcggct gcctgctagc ctgcgacgcg
     2041 caggcggtgc ggatcacgcg cattacggaa aatgccggcg cgttctttgg acgcgaaact
     2101 ccgcgggtcg gtgagctact cgccgattac ttcggcgaga ccgaagccca tgcgctgcgc
     2161 aacgcactgg cgcagtcctc cgatccaaag cgaccggcgc tgatcttcgg ttggcgcgac
     2221 ggcctgaccg gccgcacctt cgacatctca ctgcatcgcc atgacggtac atcgatcatc
     2281 gagttcgagc ctgcggcggc cgaacaggcc gacaatccgc tgcggctgac gcggcagatc
     2341 atcgcgcgca ccaaagaact gaagtcgctc gaagagatgg ccgcacgggt gccgcgctat
     2401 ctgcaggcga tgctcggcta tcaccgcgtg atgttgtacc gcttcgcgga cgacggctcc
     2461 gggatggtga tcggcgaggc gaagcgcagc gacctcgaga gctttctcgg tcagcacttt
     2521 ccggcgtcgc tggtcccgca gcaggcgcgg ctactgtact tgaagaacgc gatccgcgtg
     2581 gtctcggatt cgcgcggcat cagcagccgg atcgtgcccg agcacgacgc ctccggcgcc
     2641 gcgctcgatc tgtcgttcgc gcacctgcgc agcatctcgc cctgccatct cgaatttctg
     2701 cggaacatgg gcgtcagcgc ctcgatgtcg ctgtcgatca tcattgacgg cacgctatgg
     2761 ggattgatca tctgtcatca ttacgagccg cgtgccgtgc cgatggcgca gcgcgtcgcg
     2821 gccaaaaggt tcgccgagcg cttatcgacg cacttcaccg ccgcccacca ccaacgctag
     2881 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2941 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3001 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3061 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3121 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3181 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3241 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3301 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3361 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3421 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3481 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3541 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3601 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3661 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3721 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3841 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3901 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3961 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4021 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     4081 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4141 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4201 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4261 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     4321 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4381 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4441 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4501 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4561 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4621 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4681 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4741 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4801 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4861 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4921 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4981 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5041 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     5101 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     5161 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5221 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5281 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5341 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5401 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5461 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5521 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5581 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5641 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5701 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5761 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5821 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5881 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5941 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6001 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6061 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6121 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6181 tgattctgt
//
