LOCUS       sequence-388152-        6009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the fusion protein of miRFP720-Actin-C-18 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6009)
  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 6009)
  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: pmiRFP720-Actin-C-18
FEATURES             Location/Qualifiers
     source          1..6009
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1560
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     polyA_signal    2869..2990
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2906..2925)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2960..2979
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2997..3452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3134..3155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3346..3365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3479..3583
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3608..3628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3652..3862
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3674..3870
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3721..3856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3905..4699
                     /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             3905..4699
                     /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(3959..3978)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4569..4588
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4887..4906)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4931..4978
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5307..5895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5796..5815
                     /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 ccggtcgcca ccatggcgga aggatccgtc gccaggcagc ctgacctctt gacctgcgac
      661 gatgagccga tccatatccc cggtgccatc caaccgcatg gactgctgct cgccctcgcc
      721 gccgacatga cgatcgttgc cggcagcgac aaccttcccg aactcaccgg actggcgatc
      781 ggcgccctga tcggccgctc tgcggccgat gtcttcgact cggagacgca caaccgtctg
      841 acgatcgcct tggccgagcc cggggcggcc gtcggagcac cgatcactgt cggcttcacg
      901 atgcgaaagg acgcaggctt catcggctcc tggcatcgcc atgatcagct catcttcctc
      961 gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg cgttcttccg ccgcaccaac
     1021 agcgccatcc gccgcctgca ggccgccgaa accttggaaa gcgcctgcgc cgccgcggcg
     1081 caagaggtgc ggaagattac cggcttcgat cgggtgatga tctatcgctt cgcctccgac
     1141 ttcagcgggt ccgtgatcgc agaggatcgg tgcgccgagg tcgagtcaaa actaggcctg
     1201 cactatcctg cctcattcat cccggcgcag gcccgtcggc tctataccat caacccggta
     1261 cggatcattc ccgatatcaa ttatcggccg gtgccggtca ccccagacct caatccggtc
     1321 accgggcggc cgattgatct tagcttcgcc atcctgcgca gcgtctcgcc caaccatctg
     1381 gagttcatgc gcaacatagg catgcacggc acgatgtcga tctcgatttt gcgcggcgag
     1441 cgactgtggg gattgatcgt ttgccatcac cgaacgccgt actacgtcga tctcgatggc
     1501 cgccaagcct gcaagagagt cgccgagaga ctggccaccc agatcggcgt gatggaagag
     1561 tccggactca gatctggcag cggtggaggc agcgcatccg gcggaagcgg aagcgatgat
     1621 gatatcgccg cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg cttcgcgggc
     1681 gacgatgccc cccgggccgt cttcccctcc atcgtggggc gccccaggca ccagggcgtg
     1741 atggtgggca tgggtcagaa ggattcctat gtgggcgacg aggcccagag caagagaggc
     1801 atcctcaccc tgaagtaccc catcgagcac ggcatcgtca ccaactggga cgacatggag
     1861 aaaatctggc accacacctt ctacaatgag ctgcgtgtgg ctcccgagga gcaccccgtg
     1921 ctgctgaccg aggcccccct gaaccccaag gccaaccgcg agaagatgac ccagatcatg
     1981 tttgagacct tcaacacccc agccatgtac gttgctatcc aggctgtgct atccctgtac
     2041 gcctctggcc gtaccactgg catcgtgatg gactccggtg acggggtcac ccacactgtg
     2101 cccatctacg aggggtatgc cctcccccat gccatcctgc gtctggacct ggctggccgg
     2161 gacctgactg actacctcat gaagatcctc accgagcgcg gctacagctt caccaccacg
     2221 gccgagcggg aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc cctggacttc
     2281 gagcaagaga tggccacggc tgcttccagc tcctccctgg agaagagcta cgagctgcct
     2341 gacggccagg tcatcaccat tggcaatgag cggttccgct gccctgaggc actcttccag
     2401 ccttccttcc tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa ctccatcatg
     2461 aagtgtgacg tggacatccg caaagacctg tacgccaaca cagtgctgtc tggcggcacc
     2521 accatgtacc ctggcattgc cgacaggatg cagaaggaga tcactgccct ggcacccagc
     2581 acaatgaaga tcaagatcat tgctcctcct gagcgcaagt actccgtgtg gatcggcggc
     2641 tccatcctgg cctcgctgtc caccttccag cagatgtgga tcagcaagca ggagtatgac
     2701 gagtccggcc cctccatcgt ccaccgcaaa tgcttctagg gatccaccgg atctagataa
     2761 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2821 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3001 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3061 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3121 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3181 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3241 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3301 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3361 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3421 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3601 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3661 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3721 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3781 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3841 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3901 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3961 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4021 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4081 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4141 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4201 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4261 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4321 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4381 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4441 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4501 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4561 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4621 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4681 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4741 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4801 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4861 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4921 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4981 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5041 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5101 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5161 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5221 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6001 gccatgcat
//
