LOCUS       sequence-410749-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LC-Myosin labelled with near-infrared fluorescent protein 
            miRFP704nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Oliinyk OS, Pletnev S, Baloban M, Verkhusha VV
  TITLE     Development of bright red-shifted miRFP704nano using structural 
            analysis of miRFPnano proteins.
  JOURNAL   Protein Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709.
  PUBMED    37347539
REFERENCE   2  (bases 1 to 5993)
  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: "Protein 
            Sci. 2023 Jun 22:e4709. doi: 10.1002/pro.4709."
COMMENT     Sequence Label: pMyosin-miRFP704nano
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    19..40
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(157..174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(328..916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(408..427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1087..1878,1879..1947))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1710..1729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1948..2052)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2127..2146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2318..2697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2698..2901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2852..2872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2946..2965
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2946..2964
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3999..4028
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4044..4061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4084..4101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4090..4314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4360..4788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4447..4466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4657..4678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4797..4817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4802..5131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4982..5117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5044..5063
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5179..5226
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5245..5619
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5749..5870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5786..5805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5840..5859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5919..5935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5919..5935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5932..5954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5943..5959
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5967..5973,5974..5991,5992..5993,1..4))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
       61 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      121 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      181 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      241 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1081 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1141 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1201 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1261 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1321 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1381 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1441 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1501 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1561 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1621 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1681 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1741 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1801 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1861 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1921 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1981 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2041 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     2101 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2161 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     2221 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     2281 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     2341 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2401 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2461 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2521 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2581 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2641 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2701 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2761 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     2821 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2881 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
     2941 gaaattaata cgactcacta tagggagacc caagctggct agttaagctt ggtaccatgg
     3001 caaacctgga caagatgctg aacaccatcg tgaccgaggt gcgccagttc ctgcaagtgg
     3061 acagagtgtg cgtgttccag ttcgaggaag attactccgg cagcgtcgtc gtggaagccg
     3121 tggacgacag atggaacagc atcctgaaga cccaggtgcg ggactgctac ttcatggaaa
     3181 ccagaggtga ggaatacttg cacggcagat accaggccat cgccgacatc taccaggcca
     3241 atctgctgga gagctacaga gacctgctgg gccagtttca ggtgcgggcc attctggctg
     3301 tccccatcat caagggcaag aagctgtggg gcctgctggt ggcccaccaa ctggctgccc
     3361 ctcggagctg gcagacctgg gaaatcgagt tcctgaaaca gcaagccgtg gtgatgggca
     3421 tcgccatcca gcagagcgga ggtggtggat caggaggaag atctatgtcg agcaagagag
     3481 ccaaggccaa gaccaccaag aagaggcccc agagggctac gtccaatgtc ttcgccatgt
     3541 ttgaccagtc ccagatccag gagtttaagg aggccttcaa catgattgat cagaaccgcg
     3601 atggcttcat tgataaggag gacctgcacg acatgttggc ctctctgggg aagaacccca
     3661 cagacgagta cctggagggc atgatgaacg aggcgccagg gcccatcaac ttcaccatgt
     3721 tcctcacaat gtttggggag aagctgaacg gcacagaccc cgaggatgtg atccgcaatg
     3781 cctttgcctg ctttgatgag gaggcctcag gcttcatcca cgaggaccac ctgagggagc
     3841 tgctcaccac catgggcgac cgattcacgg atgaggaggt ggacgagatg taccgcgagg
     3901 cacccattga taagaagggc aacttcaact acgtggagtt cactcgcatc ctcaaacacg
     3961 gcgccaagga caaggacgac taatctagag ggcccttcga acaaaaactc atctcagaag
     4021 aggatctgaa tatgcatacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     4081 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4141 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4201 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4261 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4321 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4381 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4441 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4501 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4561 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4621 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4681 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4741 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4801 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4921 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4981 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5041 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5101 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5161 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     5221 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     5281 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     5341 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     5401 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     5461 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     5521 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     5581 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     5641 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5701 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5761 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5821 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5881 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5941 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtg
//
