LOCUS       sequence-360259-        5079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for iRFP670-PUFc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5079)
  AUTHORS   Clow PA, Du M, Jillette N, Taghbalout A, Zhu JJ, Cheng AW
  TITLE     CRISPR-mediated multiplexed live cell imaging of nonrepetitive 
            genomic loci with one guide RNA per locus.
  JOURNAL   Nat Commun. 2022 Apr 6;13(1):1871. doi: 10.1038/s41467-022-29343-z.
  PUBMED    35387989
REFERENCE   2  (bases 1 to 5079)
  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 
            Commun."; date: "2022-04-6"; volume: "13(1):1871. doi"; pages: " 
            10.1038/s41467-022-29343-z"
COMMENT     Sequence Label: pAC11005-pmax-iRFP670_PUFc
FEATURES             Location/Qualifiers
     source          1..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    960..984
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1008..1940
                     /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
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             2016..2036
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(3108..3132)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3185..3226
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3252..3271)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3281..3362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3306..3325
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3560..4148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3640..3659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4229..5023)
                     /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             complement(4229..5023)
                     /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(4340..4359)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4950..4969
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagg
      721 tcgtttagtg aaccgtcaga tcactagtag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctc gactgatcac aggtaagtat caaggttaca agacaggttt
      841 aaggaggcca atagaaactg ggcttgtcga gacagagaag attcttgcgt ttctgatagg
      901 cacctattgg tcttactgac atccactttg cctttctctc cacagggaaa aaacaattga
      961 caagtttgta caaaaaagca ggctccgaat tcaccggtgc cgccaccatg gcgcgtaagg
     1021 tcgatctcac ctcctgcgat cgcgagccga tccacatccc cggcagcatt cagccgtgcg
     1081 gctgcctgct agcctgcgac gcgcaggcgg tgcggatcac gcgcattacg gaaaatgccg
     1141 gcgcgttctt tggacgcgaa actccgcggg tcggtgagct actcgccgat tacttcggcg
     1201 agaccgaagc ccatgcgctg cgcaacgcac tggcgcagtc ctccgatcca aagcgaccgg
     1261 cgctgatctt cggttggcgc gacggcctga ccggccgcac cttcgacatc tcactgcatc
     1321 gccatgacgg tacatcgatc atcgagttcg agcctgcggc ggccgaacag gccgacaatc
     1381 cgctgcggct gacgcggcag atcatcgcgc gcaccaaaga actgaagtcg ctcgaagaga
     1441 tggccgcacg ggtgccgcgc tatctgcagg cgatgctcgg ctatcaccgc gtgatgttgt
     1501 accgcttcgc ggacgacggc tccgggatgg tgatcggcga ggcgaagcgc agcgacctgg
     1561 agagctttct cggtcagcac tttccggcgt cgctggtccc gcagcaggcg cggctactgt
     1621 acttgaagaa cgcgatccgc gtggtctcgg attcgcgcgg catcagcagc cggatcgtgc
     1681 ccgagcacga cgcctccggc gccgcgctcg atctgtcgtt cgcgcacctg cgcagcatct
     1741 cgccctgcca tctcgaattt ctgcggaaca tgggcgtcag cgcctcgatg tcgctgtcga
     1801 tcatcattga cggcacgcta tggggattga tcatctgtca tcattacgag ccgcgtgccg
     1861 tgccgatggc gcagcgcgtc gcggccgaaa tgttcgccga cttcttatcg ctgcacttca
     1921 ccgccgccca ccaccaacgc ggcggccgcg ggggaggcgg ttccggtggc ggcggaagcg
     1981 gaggtggagg atcagggccg gccgggatcc tcccccccaa gaaaaagagg aaggtatcta
     2041 gaggccgcag ccgccttttg gaagattttc gaaacaaccg gtaccccaat ttacaactgc
     2101 gggagattgc tggacatata atggaatttt cccaagacca gcatgggtcc agattcattc
     2161 agctgaaact ggagcgtgcc acaccagctg agcgccagct tgtcttcaat gaaatcctcc
     2221 aggctgccta ccaactcatg gtggatgtgt ttggtaatta cgtcattcag aagttctttg
     2281 aatttggcag tcttgaacag aagctggctt tggcagaacg gattcgaggc cacgtcctgt
     2341 cattggcact acagatgtat ggcagccgtg ttatcgagaa agctcttgag tttattcctt
     2401 cagaccagca gaatgagatg gttcgggaac tagatggcca tgtcttgaag tgtgtgaaag
     2461 atcagaatgg caatcacgtg gttcagaaat gcattgaatg tgtacagccc cagtctttgc
     2521 aatttatcat cgacgcgttt aagggacagg tatttgcctt atccacacat ccttatggct
     2581 gccgagtgat tcagagaatc ctggagcact gtctccctga ccagacactc cctattttag
     2641 aggagcttca ccagcacaca gagcagctgg tacaggatca atatggaagt tatgtaatcg
     2701 aacatgtact ggagcacggt cgtcctgagg ataaaagcaa aattgtagca gaaatccgag
     2761 gcaatgtact tgtattgagt cagcacaaat ttgcaaacaa tgttgtgcag aagtgtgtta
     2821 ctcacgcctc acgtacggag cgcgctgtgc tcatcgacga ggtgtgcacc atgaacgacg
     2881 gtccccacag tgccttatac accatgatga aggaccagta tgccaactac gtggtccaga
     2941 agatgattga cgtggcggag ccaggccagc ggaagatcgt catgcataag atccggcccc
     3001 acatcgcaac tcttcgtaag tacacctatg gcaagcacat tctggccaag ctggagaagt
     3061 actacatgaa gaacggtgtt gacttagggt gattaattaa gaattcgacc cagctttctt
     3121 gtacaaagtg gttgatatcc agcacagtgg cggccgctcg agtctagagg gcccgcggtt
     3181 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggttagca
     3241 attgtttttt cgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3301 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3361 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3421 tttaaagcaa gtaaaacctc tacaaatgtg gtacttaaga gggggagacc aaagggcgag
     3481 acgttaaggc ctcacgtgac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3541 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3601 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3661 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttacggga tacctgtccg
     3721 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3781 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3841 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3901 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3961 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4021 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4081 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4141 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4201 cacgttaagg gattttggtc atgccgtctc agaagaactc gtcaagaagg cgatagaagg
     4261 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     4321 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     4381 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     4441 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     4501 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     4561 gatcgacaag accggcttcc atccgagtac gtgctctctc gatgcgatgt ttcgcttggt
     4621 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     4681 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     4741 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagtacagct gcgcaaggaa
     4801 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     4861 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     4921 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     4981 aagcggccgg agaacctgcg tgcaatccat cttgttcaat cataatatta ttgaagcatt
     5041 tatcagggtt cgtctcgtcc cggtctcctc ccatgcatg
//
