LOCUS       sequence-410836-        6722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human beta-actin labelled with near-infrared fluorescent protein 
            miRFP704nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  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 6722)
  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: pActin-miRFP704nano
FEATURES             Location/Qualifiers
     source          1..6722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(60..648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(140..159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(819..1610,1611..1679))
                     /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     1442..1461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1680..1784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1859..1878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2050..2429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2430..2633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2584..2604
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2678..2697
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2678..2696
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4460..4489
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4505..4522
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4545..4562)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4551..4775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4821..5249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4908..4927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5118..5139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5258..5278)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5263..5592
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5443..5578
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5505..5524
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5640..5687
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5706..6080
                     /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    6210..6331
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6247..6266)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6301..6320
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6380..6396)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6380..6396)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6393..6415)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6404..6420
                     /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(6428..6434,6435..6452,6453..6458))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6473..6494
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6611..6628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
       61 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
      841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
      901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
      961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     1141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     1441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1801 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     1861 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     1921 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     1981 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     2041 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     2101 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2161 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2221 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2281 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2341 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2401 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2461 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2521 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2581 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     2641 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     2701 cccaagctgg ctagttaagc ttggtaccat ggcaaacctg gacaagatgc tgaacaccat
     2761 cgtgaccgag gtgcgccagt tcctgcaagt ggacagagtg tgcgtgttcc agttcgagga
     2821 agattactcc ggcagcgtcg tcgtggaagc cgtggacgac agatggaaca gcatcctgaa
     2881 gacccaggtg cgggactgct acttcatgga aaccagaggt gaggaatact tgcacggcag
     2941 ataccaggcc atcgccgaca tctaccaggc caatctgctg gagagctaca gagacctgct
     3001 gggccagttt caggtgcggg ccattctggc tgtccccatc atcaagggca agaagctgtg
     3061 gggcctgctg gtggcccacc aactggctgc ccctcggagc tggcagacct gggaaatcga
     3121 gttcctgaaa cagcaagccg tggtgatggg catcgccatc cagcagagcg gaggtggtgg
     3181 atcaggagga ggtggttctg gaggtggtgg atcaggagga ggtggttctg gaggtggtgg
     3241 atcaggagga ggtggttctg gaggtggtgg atcaggagga ggtggttctg gaggtggtgg
     3301 atcaggagga agatctatgg atgatgatat cgccgcgctc gtcgtcgaca acggctccgg
     3361 catgtgcaag gccggcttcg cgggcgacga tgccccccgg gccgtcttcc cctccatcgt
     3421 ggggcgcccc aggcaccagg gcgtgatggt gggcatgggt cagaaggatt cctatgtggg
     3481 cgacgaggcc cagagcaaga gaggcatcct caccctgaag taccccatcg agcacggcat
     3541 cgtcaccaac tgggacgaca tggagaaaat ctggcaccac accttctaca atgagctgcg
     3601 tgtggctccc gaggagcacc ccgtgctgct gaccgaggcc cccctgaacc ccaaggccaa
     3661 ccgcgagaag atgacccaga tcatgtttga gaccttcaac accccagcca tgtacgttgc
     3721 tatccaggct gtgctatccc tgtacgcctc tggccgtacc actggcatcg tgatggactc
     3781 cggtgacggg gtcacccaca ctgtgcccat ctacgagggg tatgccctcc cccatgccat
     3841 cctgcgtctg gacctggctg gccgggacct gactgactac ctcatgaaga tcctcaccga
     3901 gcgcggctac agcttcacca ccacggccga gcgggaaatc gtgcgtgaca ttaaggagaa
     3961 gctgtgctac gtcgccctgg acttcgagca agagatggcc acggctgctt ccagctcctc
     4021 cctggagaag agctacgagc tgcctgacgg ccaggtcatc accattggca atgagcggtt
     4081 ccgctgccct gaggcactct tccagccttc cttcctgggc atggagtcct gtggcatcca
     4141 cgaaactacc ttcaactcca tcatgaagtg tgacgtggac atccgcaaag acctgtacgc
     4201 caacacagtg ctgtctggcg gcaccaccat gtaccctggc attgccgaca ggatgcagaa
     4261 ggagatcact gccctggcac ccagcacaat gaagatcaag atcattgctc ctcctgagcg
     4321 caagtactcc gtgtggatcg gcggctccat cctggcctcg ctgtccacct tccagcagat
     4381 gtggatcagc aagcaggagt atgacgagtc cggcccctcc atcgtccacc gcaaatgctt
     4441 ctagtctaga gggcccttcg aacaaaaact catctcagaa gaggatctga atatgcatac
     4501 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     4561 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4621 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4681 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     4741 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4801 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4861 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4921 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4981 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5041 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5101 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5161 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5221 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     5281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5521 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5581 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     5641 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     5701 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     5761 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     5821 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     5881 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     5941 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     6001 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     6061 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     6121 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6181 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6241 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6301 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6361 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6421 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6481 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6541 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6721 ac
//
