LOCUS       sequence-410752-        5491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Near-infrared fluorescent protein miRFP704nano in mammalian plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5491)
  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 5491)
  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: pcDNA-miRFP704nano
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1568..1587)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1759..2138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2139..2342
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2293..2313
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2387..2406
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2387..2405
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2938..2967
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2983..3000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3023..3040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3029..3253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3299..3727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3386..3405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3596..3617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3736..3756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3741..4070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3921..4056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3983..4002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4118..4165
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4184..4558
                     /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    4688..4809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4725..4744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4779..4798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4858..4874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4858..4874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4871..4893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4882..4898
                     /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(4906..4912,4913..4930,4931..4936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4951..4972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5089..5106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5260..5491,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5340..5359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     1561 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     1621 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     1681 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     1741 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1801 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1861 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1921 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1981 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2041 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2101 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2161 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2221 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2281 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2341 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     2401 atagggagac ccaagctggc tagttaagct tggtaccatg gcaaacctgg acaagatgct
     2461 gaacaccatc gtgaccgagg tgcgccagtt cctgcaagtg gacagagtgt gcgtgttcca
     2521 gttcgaggaa gattactccg gcagcgtcgt cgtggaagcc gtggacgaca gatggaacag
     2581 catcctgaag acccaggtgc gggactgcta cttcatggaa accagaggtg aggaatactt
     2641 gcacggcaga taccaggcca tcgccgacat ctaccaggcc aatctgctgg agagctacag
     2701 agacctgctg ggccagtttc aggtgcgggc cattctggct gtccccatca tcaagggcaa
     2761 gaagctgtgg ggcctgctgg tggcccacca actggctgcc cctcggagct ggcagacctg
     2821 ggaaatcgag ttcctgaaac agcaagccgt ggtgatgggc atcgccatcc agcagagcta
     2881 gctgaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccttcgaa
     2941 caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat
     3001 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3061 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3121 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3181 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3241 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3301 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3361 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3421 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3481 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3541 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3601 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3661 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3721 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3841 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3901 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3961 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4021 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4081 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4141 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     4201 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     4261 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     4321 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     4381 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     4441 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     4501 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     4561 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4681 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4801 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5461 acgctgtagg tatctcagtt cggtgtaggt c
//
