LOCUS       sequence-364932-        6023 bp    DNA     circular SYN 14-APR-2025
DEFINITION  LC-Myosin labelled with near-infrared fluorescent protein 
            miRFP670nano3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  AUTHORS   Oliinyk OS, Baloban M, Clark CL, Carey E, Pletnev S, Nimmerjahn A, 
            Verkhusha VV
  TITLE     Single-domain near-infrared protein provides a scaffold for 
            antigen-dependent fluorescent nanobodies.
  JOURNAL   Nat Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6.
  PUBMED    35606446
REFERENCE   2  (bases 1 to 6023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6."
COMMENT     Sequence Label: pMyosin-miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1946..1975
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1991..2008
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2031..2048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2037..2261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2307..2735
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2394..2413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2604..2625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2744..2764)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2749..3078
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2929..3064
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2991..3010
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3126..3173
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3192..3566
                     /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    3696..3817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3733..3752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3787..3806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3866..3882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3866..3882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3879..3901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3890..3906
                     /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(3914..3920,3921..3938,3939..3944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3959..3980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4097..4114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4268..4856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4348..4367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5027..5818,5819..5887))
                     /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     5650..5669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5888..5992)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accatggcaa acctggacaa gatgctgaac accaccgtga ccgaggtgcg
      961 caagttcctg caagcggaca gagtgtgcgt gttcaagttc gaggaagatt actccggcac
     1021 cgtctcgcac gaagccgtgg acgacagatg gattagcatc ctgaagaccc aggtgcagga
     1081 cagatacttc atggaaacca gaggcgagga atacgtccac ggcagatacc aggccatcgc
     1141 cgacatctac acagccaatc tggtcgagtg ctacagagac ctgctgatcg agtttcaggt
     1201 gcgggccatt ctggctgtcc ccatcctgca aggcaagaag ctgtggggcc tgctggtggc
     1261 ccaccaactg gccggccctc gggagtggca gacctgggaa atcgacttcc tgaaacagca
     1321 agccgtggtg atgggcatcg ccatccagca gagcggaggt ggtggatcag gaggaggtgg
     1381 ttctggaggt ggtggatcag gaggaagatc tatgtcgagc aagagagcca aggccaagac
     1441 caccaagaag aggccccaga gggctacgtc caatgtcttc gccatgtttg accagtccca
     1501 gatccaggag tttaaggagg ccttcaacat gattgatcag aaccgcgatg gcttcattga
     1561 taaggaggac ctgcacgaca tgttggcctc tctggggaag aaccccacag acgagtacct
     1621 ggagggcatg atgaacgagg cgccagggcc catcaacttc accatgttcc tcacaatgtt
     1681 tggggagaag ctgaacggca cagaccccga ggatgtgatc cgcaatgcct ttgcctgctt
     1741 tgatgaggag gcctcaggct tcatccacga ggaccacctg agggagctgc tcaccaccat
     1801 gggcgaccga ttcacggatg aggaggtgga cgagatgtac cgcgaggcac ccattgataa
     1861 gaagggcaac ttcaactacg tggagttcac tcgcatcctc aaacacggcg ccaaggacaa
     1921 ggacgactaa tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat
     1981 gcataccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     2041 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2101 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2161 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2221 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2281 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2341 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2401 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2461 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2521 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2581 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2641 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2701 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2761 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2821 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2881 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2941 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3001 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3061 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     3121 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3181 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3241 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3301 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3361 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     3421 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     3481 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     3541 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     3601 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3661 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3841 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4081 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4681 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6001 cccgaaaagt gccacctgac gtc
//
