LOCUS       sequence-365042-        6632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human beta-actin labelled with near-infrared fluorescent protein 
            miRFP670nano3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6632)
  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 6632)
  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. 2022 May 23. pii: 10.1038/s41592-022-01467-6. doi: 
            10.1038/s41592-022-01467-6."
COMMENT     Sequence Label: pActin-miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..6632
                     /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             2555..2584
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2600..2617
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2640..2657)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2646..2870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2916..3344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3003..3022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3213..3234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3353..3373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3358..3687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3538..3673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3600..3619
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3735..3782
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3801..4175
                     /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    4305..4426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4342..4361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4396..4415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4475..4491)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4475..4491)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4488..4510)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4499..4515
                     /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(4523..4529,4530..4547,4548..4553))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4568..4589
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4706..4723)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4877..5465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4957..4976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5636..6427,6428..6496))
                     /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     6259..6278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6497..6601)
                     /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 tatggatgat gatatcgccg cgctcgtcgt
     1441 cgacaacggc tccggcatgt gcaaggccgg cttcgcgggc gacgatgccc cccgggccgt
     1501 cttcccctcc atcgtggggc gccccaggca ccagggcgtg atggtgggca tgggtcagaa
     1561 ggattcctat gtgggcgacg aggcccagag caagagaggc atcctcaccc tgaagtaccc
     1621 catcgagcac ggcatcgtca ccaactggga cgacatggag aaaatctggc accacacctt
     1681 ctacaatgag ctgcgtgtgg ctcccgagga gcaccccgtg ctgctgaccg aggcccccct
     1741 gaaccccaag gccaaccgcg agaagatgac ccagatcatg tttgagacct tcaacacccc
     1801 agccatgtac gttgctatcc aggctgtgct atccctgtac gcctctggcc gtaccactgg
     1861 catcgtgatg gactccggtg acggggtcac ccacactgtg cccatctacg aggggtatgc
     1921 cctcccccat gccatcctgc gtctggacct ggctggccgg gacctgactg actacctcat
     1981 gaagatcctc accgagcgcg gctacagctt caccaccacg gccgagcggg aaatcgtgcg
     2041 tgacattaag gagaagctgt gctacgtcgc cctggacttc gagcaagaga tggccacggc
     2101 tgcttccagc tcctccctgg agaagagcta cgagctgcct gacggccagg tcatcaccat
     2161 tggcaatgag cggttccgct gccctgaggc actcttccag ccttccttcc tgggcatgga
     2221 gtcctgtggc atccacgaaa ctaccttcaa ctccatcatg aagtgtgacg tggacatccg
     2281 caaagacctg tacgccaaca cagtgctgtc tggcggcacc accatgtacc ctggcattgc
     2341 cgacaggatg cagaaggaga tcactgccct ggcacccagc acaatgaaga tcaagatcat
     2401 tgctcctcct gagcgcaagt actccgtgtg gatcggcggc tccatcctgg cctcgctgtc
     2461 caccttccag cagatgtgga tcagcaagca ggagtatgac gagtccggcc cctccatcgt
     2521 ccaccgcaaa tgcttctagt ctagagggcc cttcgaacaa aaactcatct cagaagagga
     2581 tctgaatatg cataccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2881 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3241 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3601 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3661 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3721 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     3781 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     3841 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     3901 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     3961 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     4021 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     4081 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     4141 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     4201 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4261 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     4321 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4381 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4441 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4501 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4561 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4621 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4681 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4741 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4801 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4861 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5281 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5341 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5401 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5461 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5521 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5581 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5701 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5821 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6061 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6301 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6481 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6601 gcacatttcc ccgaaaagtg ccacctgacg tc
//
