LOCUS       sequence-208118-        4953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP in mammalian 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4953)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 4953)
  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 Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pmiRFP-N1
FEATURES             Location/Qualifiers
     source          1..4953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        770..1073
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1074..1277
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1228..1248
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1300..1380
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1381..1390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2450..2571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2487..2506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2541..2560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2578..3033)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2715..2736)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2927..2946
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3060..3164
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3166..3523
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3166..3357
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3189..3209)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3374..3509
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3558..4352
                     /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             3558..4352
                     /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(3612..3631)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4222..4241
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4540..4559)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4584..4631
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
      721 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      781 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      841 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      901 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      961 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1021 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1081 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1141 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1201 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1261 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggactcaga
     1321 tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt
     1381 gccaccatgg tcgcaggaca cgcaagtgga agccccgact ttggaaccgc cgatccgagc
     1441 gactgtgaga gagaagaaat ccatctggcc ggatccatcc agccacacgg aacactgctg
     1501 gtggtctccg agcctgacca tagaatcatt caggcatctg ccaacgccgc tgaattcctg
     1561 aatctgggaa gtgtgctggg agtccccctg gctgagatcg acggggatct gctgatcaag
     1621 attctgcccc acctggaccc taccgcagaa ggaatgccag tggccgtcag gtgccgcatt
     1681 ggaaacccct ccacagagta cgatggactg atgcatcgac cacctgaagg aggactgatc
     1741 attgagctgg aacgggctgg gccacccatt gatctgagcg gaacactggc tcccgcactg
     1801 gagcgaattc gaactgcagg atccctgagg gctctgtgcg acgatactgc actgctgttc
     1861 cagcagtgta ccggctacga ccgagtgatg gtctatcggt tcgatgaaca ggggcacgga
     1921 gaggtgtatt cagaaataca tgtgactgga ctggagagct actttggcaa caggtatcct
     1981 agctccctcg tgccacagat ggcccggaga ctgtacgaaa gacagagggt gcgcgtcctg
     2041 gtggatgtct cttatcagcc tgtgccactg gagccaagac tgagtcccct gactggcagg
     2101 gacctggata tgtcagggtg ttttctgcgc tctatgagtc ctactcacct gcagttcctg
     2161 aagaatatgg gcgtgcgggc aacactggtg gtcagcctgg tggtcggagg caaactgtgg
     2221 gggctggtga tctgccacca ttatctgcca cgcttcatcc attttgagct gcgagcaatt
     2281 tgtgagctgc tggccgaagc tatcgcaacc cgcattacag ccctgtaagc ggccgcgact
     2341 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2401 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2461 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2521 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     2581 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2641 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2701 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2761 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2821 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2881 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2941 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3001 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3061 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3181 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3241 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3301 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3361 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3421 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3481 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     3541 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3601 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3661 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3721 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3781 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3841 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3901 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3961 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4021 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4081 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4141 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4201 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4261 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4321 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4381 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4441 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4501 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     4561 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     4621 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     4681 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     4741 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     4801 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4861 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4921 ctgagcgtca gaccccgtag aaaagatcaa agg
//
