LOCUS       sequence-150054-        7602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha actinin labeling with near-infrared fluorescent protein 
            miRFP703.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  AUTHORS   Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, 
            Verkhusha VV
  TITLE     Bright monomeric near-infrared fluorescent proteins as tags and 
            biosensors for multiscale imaging.
  JOURNAL   Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
  PUBMED    27539380
REFERENCE   2  (bases 1 to 7602)
  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: "2016-08-19"; volume: "7:12405. doi"; pages: " 
            10.1038/ncomms12405"
COMMENT     Sequence Label: pActinin-miRFP703
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3364..3373
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3370..4317
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 703 nm (Shcherbakova et al., 2016)"
                     /label=miRFP703
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSLVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPIHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     polyA_signal    4439..4560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4476..4495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4530..4549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4567..5022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4704..4725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4916..4935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5155..5512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5155..5346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5178..5198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5363..5498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5547..6341
                     /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             5547..6341
                     /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(5601..5620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6211..6230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6529..6548)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6573..6620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6949..7537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7438..7457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcgc accatcatgg accattatga ttctcagcaa
      661 accaacgatt acatgcagcc agaagaggac tgggaccggg acctgctcct ggacccggcc
      721 tgggagaagc agcagagaaa gacattcacg gcatggtgta actcccacct ccggaaggcg
      781 gggacacaga tcgagaacat cgaagaggac ttccgggatg gcctgaagct catgctgctg
      841 ctggaggtca tctcaggtga acgcttggcc aagccagagc gaggcaagat gagagtgcac
      901 aagatctcca acgtcaacaa ggccctggat ttcatagcca gcaaaggcgt caaactggtg
      961 tccatcggag ccgaagaaat cgtggatggg aatgtgaaga tgaccctggg catgatctgg
     1021 accatcatcc tgcgctttgc catccaggac atctccgtgg aagagacttc agccaaggaa
     1081 gggctgctcc tgtggtgtca gagaaagaca gccccttaca aaaatgtcaa catccagaac
     1141 ttccacataa gctggaagga tggcctcggc ttctgtgctt tgatccaccg acaccggccc
     1201 gagctgattg actacgggaa gctgcggaag gatgatccac tcacaaatct gaatacggct
     1261 tttgacgtgg cagagaagta cctggacatc cccaagatgc tggatgccga agacatcgtt
     1321 ggaactgccc gaccggatga gaaagccatc atgacttacg tgtctagctt ctaccacgcc
     1381 ttctctggag cccagaaggc ggagacagca gccaatcgca tctgcaaggt gttggccgtc
     1441 aaccaggaga acgagcagct tatggaagac tacgagaagc tggccagtga tctgttggag
     1501 tggatccgcc gcacaatccc gtggctggag aaccgggtgc ccgagaacac catgcatgcc
     1561 atgcaacaga agctggagga cttccgggac taccggcgcc tgcacaagcc gcccaaggtg
     1621 caggagaagt gccagctgga gatcaacttc aacacgctgc agaccaagct gcggctcagc
     1681 aaccggcctg ccttcatgcc ctctgagggc aggatggtct cggacatcaa caatgcctgg
     1741 ggctgcctgg agcaggtgga gaagggctat gaggagtggt tgctgaatga gatccggagg
     1801 ctggagcgac tggaccacct ggcagagaag ttccggcaga aggcctccat ccacgaggcc
     1861 tggactgacg gcaaagaggc catgctgcga cagaaggact atgagaccgc caccctctcg
     1921 gagatcaagg ccttgctcaa gaagcatgag gccttcgaga gtgacctggc tgcccaccag
     1981 gaccgtgtgg agcagattgc cgccatcgca caggagctca atgagctgga ctattatgac
     2041 tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc agtgggacaa tctgggggcc
     2101 ctaactcaga agcgaaggga agctctggag cggaccgaga aactgctgga gaccattgac
     2161 cagctgtact tggagtatgc caagcgggct gcacccttca acaactggat ggagggggcc
     2221 atggaggacc tgcaggacac cttcattgtg cacaccattg aggagatcca gggactgacc
     2281 acagcccatg agcagttcaa ggccaccctc cctgatgccg acaaggagcg cctggccatc
     2341 ctgggcatcc acaatgaggt gtccaagatt gtccagacct accacgtcaa tatggcgggc
     2401 accaacccct acacaaccat cacgcctcag gagatcaatg gcaaatggga ccacgtgcgg
     2461 cagctggtgc ctcggaggga ccaagctctg acggaggagc atgcccgaca gcagcacaat
     2521 gagaggctac gcaagcagtt tggagcccag gccaatgtca tcgggccctg gatccagacc
     2581 aagatggagg agatcgggag gatctccatt gagatgcatg ggaccctgga ggaccagctc
     2641 agccacctgc ggcagtatga gaagagcatc gtcaactaca agccaaagat tgatcagctg
     2701 gagggcgacc accagctcat ccaggaggcg ctcatcttcg acaacaagca caccaactac
     2761 accatggagc acatccgtgt gggctgggag cagctgctca ccaccatcgc caggaccatc
     2821 aatgaggtag agaaccagat cctgacccgg gatgccaagg gcatcagcca ggagcagatg
     2881 aatgagttcc gggcctcctt caaccacttt gaccgggatc actccggcac actgggtccc
     2941 gaggagttca aagcctgcct catcagcttg ggttatgata ttggcaacga cccccaggga
     3001 gaagcagaat ttgcccgcat catgagcatt gtggacccca accgcctggg ggtagtgaca
     3061 ttccaggcct tcattgactt catgtcccgc gagacagccg acacagatac agcagaccaa
     3121 gtcatggctt ccttcaagat cctggctggg gacaagaact acattaccat ggacgagctg
     3181 cgccgcgagc tgccacccga ccaggctgag tactgcatcg cgcggatggc cccctacacc
     3241 ggccccgact ccgtgccagg tgctctggac tacatgtcct tctccacggc gctgtacggc
     3301 gagagtgacc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3361 gtcgccacca tggtagcagg tcatgcctct ggcagccccg cattcgggac cgcctctcat
     3421 tcgaattgcg aacatgaaga gatccacctc gccggctcga tccagccgca tggcgcgctt
     3481 ctggtcgtca gcgaacatga tcatcgcgtc atccaggcca gcgccaacgc cgcggaattt
     3541 ctgaatctcg gaagcgtact cggcgttccg ctcgccgaga tcgacggcga tctgttgatc
     3601 aagatcctgc cgcatctcga tcccaccgcc gaaggcatgc cggtcgcggt gcgctgccgg
     3661 atcggcaatc cctctacgga gtactgcggt ctgatgcatc ggcctccgga aggcgggctg
     3721 atcatcgaac tcgaacgtgc cggcccgtcg atcgatctgt caggcacgct ggcgccggcg
     3781 ctggagcgga tccgcacggc gggttcactg cgcgcgctgt gcgatgacac cgtgctgctg
     3841 tttcagcagt gcaccggcta cgaccgggtg atggtgtatc gtttcgatga gcaaggccac
     3901 ggcctggtat tctccgagtg ccatgtgcct gggctcgaat cctatttcgg caaccgctat
     3961 ccgtcgtcgc tggtcccgca gatggcgcgg cagctgtacg tgcggcagcg cgtccgcgtg
     4021 ctggtcgacg tcacctatca gccggtgccg ctggagccgc ggctgtcgcc gctgaccggg
     4081 cgcgatctcg acatgtcggg ctgcttcctg cgctcgatgt cgccgatcca tctgcagttc
     4141 ctgaaggaca tgggcgtgcg cgccaccctg gcggtgtcgc tggtggtcgg cggcaagctg
     4201 tggggcctgg ttgtctgtca ccattatctg ccgcgcttca tccgtttcga gctgcgggcg
     4261 atctgcaaac ggctcgccga aaggatcgcg acgcggatca ccgcgcttga gagctaagcg
     4321 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4381 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4561 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4621 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4681 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4741 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4801 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4861 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4921 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4981 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     5161 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     5221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     5281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5521 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5581 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5641 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5701 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5761 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5821 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5881 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5941 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6001 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6061 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6121 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6181 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6241 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6301 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6361 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6421 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6481 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6541 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6601 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6661 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6721 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6781 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7081 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7561 acggttcctg gccttttgct ggccttttgc tcacatgttc tt
//
