LOCUS       sequence-250323-        7148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha actinin labeling with near-infrared fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  AUTHORS   Oliinyk OS, Shemetov AA, Pletnev S, Shcherbakova DM, Verkhusha VV
  TITLE     Smallest near-infrared fluorescent protein evolved from 
            cyanobacteriochrome as versatile tag for spectral multiplexing.
  JOURNAL   Nat Commun. 2019 Jan 17;10(1):279. doi: 10.1038/s41467-018-08050-8.
  PUBMED    30655515
REFERENCE   2  (bases 1 to 7148)
  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: "2019-01-17"; volume: "10(1):279. doi"; pages: " 
            10.1038/s41467-018-08050-8"
COMMENT     Sequence Label: pActinin-miRFP670nano
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    3947..4068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3984..4003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4038..4057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4075..4530)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4212..4233)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4424..4443
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4663..5020
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4663..4854
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4686..4706)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4871..5006
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5055..5849
                     /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             5055..5849
                     /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(5109..5128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5719..5738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6037..6056)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6081..6128
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6457..7045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6946..6965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgagc tcaagcttcg caccatcatg gaccattatg
      661 attctcagca aaccaacgat tacatgcagc cagaagagga ctgggaccgg gacctgctcc
      721 tggacccggc ctgggagaag cagcagagaa agacattcac ggcatggtgt aactcccacc
      781 tccggaaggc ggggacacag atcgagaaca tcgaagagga cttccgggat ggcctgaagc
      841 tcatgctgct gctggaggtc atctcaggtg aacgcttggc caagccagag cgaggcaaga
      901 tgagagtgca caagatctcc aacgtcaaca aggccctgga tttcatagcc agcaaaggcg
      961 tcaaactggt gtccatcgga gccgaagaaa tcgtggatgg gaatgtgaag atgaccctgg
     1021 gcatgatctg gaccatcatc ctgcgctttg ccatccagga catctccgtg gaagagactt
     1081 cagccaagga agggctgctc ctgtggtgtc agagaaagac agccccttac aaaaatgtca
     1141 acatccagaa cttccacata agctggaagg atggcctcgg cttctgtgct ttgatccacc
     1201 gacaccggcc cgagctgatt gactacggga agctgcggaa ggatgatcca ctcacaaatc
     1261 tgaatacggc ttttgacgtg gcagagaagt acctggacat ccccaagatg ctggatgccg
     1321 aagacatcgt tggaactgcc cgaccggatg agaaagccat catgacttac gtgtctagct
     1381 tctaccacgc cttctctgga gcccagaagg cggagacagc agccaatcgc atctgcaagg
     1441 tgttggccgt caaccaggag aacgagcagc ttatggaaga ctacgagaag ctggccagtg
     1501 atctgttgga gtggatccgc cgcacaatcc cgtggctgga gaaccgggtg cccgagaaca
     1561 ccatgcatgc catgcaacag aagctggagg acttccggga ctaccggcgc ctgcacaagc
     1621 cgcccaaggt gcaggagaag tgccagctgg agatcaactt caacacgctg cagaccaagc
     1681 tgcggctcag caaccggcct gccttcatgc cctctgaggg caggatggtc tcggacatca
     1741 acaatgcctg gggctgcctg gagcaggtgg agaagggcta tgaggagtgg ttgctgaatg
     1801 agatccggag gctggagcga ctggaccacc tggcagagaa gttccggcag aaggcctcca
     1861 tccacgaggc ctggactgac ggcaaagagg ccatgctgcg acagaaggac tatgagaccg
     1921 ccaccctctc ggagatcaag gccttgctca agaagcatga ggccttcgag agtgacctgg
     1981 ctgcccacca ggaccgtgtg gagcagattg ccgccatcgc acaggagctc aatgagctgg
     2041 actattatga ctcacccagt gtcaacgccc gttgccaaaa gatctgtgac cagtgggaca
     2101 atctgggggc cctaactcag aagcgaaggg aagctctgga gcggaccgag aaactgctgg
     2161 agaccattga ccagctgtac ttggagtatg ccaagcgggc tgcacccttc aacaactgga
     2221 tggagggggc catggaggac ctgcaggaca ccttcattgt gcacaccatt gaggagatcc
     2281 agggactgac cacagcccat gagcagttca aggccaccct ccctgatgcc gacaaggagc
     2341 gcctggccat cctgggcatc cacaatgagg tgtccaagat tgtccagacc taccacgtca
     2401 atatggcggg caccaacccc tacacaacca tcacgcctca ggagatcaat ggcaaatggg
     2461 accacgtgcg gcagctggtg cctcggaggg accaagctct gacggaggag catgcccgac
     2521 agcagcacaa tgagaggcta cgcaagcagt ttggagccca ggccaatgtc atcgggccct
     2581 ggatccagac caagatggag gagatcggga ggatctccat tgagatgcat gggaccctgg
     2641 aggaccagct cagccacctg cggcagtatg agaagagcat cgtcaactac aagccaaaga
     2701 ttgatcagct ggagggcgac caccagctca tccaggaggc gctcatcttc gacaacaagc
     2761 acaccaacta caccatggag cacatccgtg tgggctggga gcagctgctc accaccatcg
     2821 ccaggaccat caatgaggta gagaaccaga tcctgacccg ggatgccaag ggcatcagcc
     2881 aggagcagat gaatgagttc cgggcctcct tcaaccactt tgaccgggat cactccggca
     2941 cactgggtcc cgaggagttc aaagcctgcc tcatcagctt gggttatgat attggcaacg
     3001 acccccaggg agaagcagaa tttgcccgca tcatgagcat tgtggacccc aaccgcctgg
     3061 gggtagtgac attccaggcc ttcattgact tcatgtcccg cgagacagcc gacacagata
     3121 cagcagacca agtcatggct tccttcaaga tcctggctgg ggacaagaac tacattacca
     3181 tggacgagct gcgccgcgag ctgccacccg accaggctga gtactgcatc gcgcggatgg
     3241 ccccctacac cggccccgac tccgtgccag gtgctctgga ctacatgtcc ttctccacgg
     3301 cgctgtacgg cgagagtgac ctcaagcttc gaattctgca gtcgacggta cccatggcaa
     3361 acctggacaa gatgctgaat accacagtaa cagaggtgcg gcagttcctg caggtggaca
     3421 gagtgtgcgt gttccagttt gaggaggatt atagcggagt ggtggtggtg gaggccgtgg
     3481 acgataggtg gatctccatc ctgaagaccc aggtgcggga tagatacttc atggagacaa
     3541 ggggcgagga gtattctcac ggccgctacc aggccatcgc cgacatctac accgcaaacc
     3601 tgacagagtg ctacagggat ctgctgacac agtttcaggt gagagcaatc ctggccgtgc
     3661 ccatcctgca gggcaagaag ctgtggggcc tgttggtggc acaccagctg gcggccccta
     3721 gacagtggca gacctgggag atcgactttc tgaagcagca ggccgtggtg gtgggcatcg
     3781 ccatccagca gagctagctg aattctgcag atatccagca cagtggcggc cgcgactcta
     3841 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3901 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     4081 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4141 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4201 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4261 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4321 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4381 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4441 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4501 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4681 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4741 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4801 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4861 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4921 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4981 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     5041 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5101 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5161 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5221 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5281 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5341 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5401 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5461 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5521 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5581 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5641 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5701 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5761 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5821 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5881 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5941 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6001 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     6061 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     6121 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     6181 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6241 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6301 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6361 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6421 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6481 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6541 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6601 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6661 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6721 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6781 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6841 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6901 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6961 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7021 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7081 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7141 ccgtatta
//
