LOCUS       sequence-383919-        5491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomeric near-infrared fluorescent protein miRFP718nano in 
            mammalian plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5491)
  AUTHORS   Oliinyk OS, Ma C, Pletnev S, Baloban M, Toboada C, Sheng H, Yao J, 
            Verkhusha VV
  TITLE     Deep-tissue SWIR imaging using rationally designed small red-shifted
            near-infrared fluorescent protein.
  JOURNAL   Nat Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0.
  PUBMED    36456785
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: "Nat 
            Methods. 2022 Dec 1. doi: 10.1038/s41592-022-01683-0."
COMMENT     Sequence Label: pcDNA-miRFP718nano
FEATURES             Location/Qualifiers
     source          1..5491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        141..520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        521..724
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     675..695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     769..788
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        769..787
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1320..1349
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1365..1382
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1405..1422)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1411..1635
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1681..2109
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1768..1787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1978..1999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2118..2138)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2123..2452
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2303..2438
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2365..2384
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2500..2547
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2566..2940
                     /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    3070..3191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3107..3126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3161..3180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3240..3256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3240..3256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3253..3275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3264..3280
                     /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(3288..3294,3295..3312,3313..3318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3333..3354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3471..3488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3642..4230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3722..3741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4401..5192,5193..5261))
                     /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     5024..5043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5262..5366)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5441..5460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
       61 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      121 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      721 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      781 ctatagggag acccaagctg gctagttaag cttggtacca tggcaaacct ggacaagatg
      841 ctgaacacca tcgtgaccga ggtgcgccag ttcctgcaag tggacagact ctgcgtgttc
      901 aagttcgagg aagattactc cggcaacatc atctacgaag ccgtggacga cggatggttg
      961 agcatcctga agacccacgt gcgggactgc tacttcatgg aaaccagagg cgaggaatac
     1021 ctgcacggca gataccaggc catcgccgac atccaccagg ccaatctggc ggagagctat
     1081 agagacttcc tgacccagta ccaggtgcgg gccattgtgg ctgtccccat cctgaagggc
     1141 aagaagctgt ggggcttgtt cagcgcccac caactggccg cccctcggag ctggcaggcc
     1201 tgggaaatcg agttcctgaa acagcaagcc gtggtgatgg gcatcgccat ccagcagagc
     1261 tagctgaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccttcg
     1321 aacaaaaact catctcagaa gaggatctga atatgcatac cggtcatcat caccatcacc
     1381 attgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1441 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1501 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1561 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1621 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     1681 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1741 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1801 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1861 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1921 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1981 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2041 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2101 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2161 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2221 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2281 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2341 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2401 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     2461 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     2521 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     2581 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     2641 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     2701 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     2761 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     2821 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     2881 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     2941 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3001 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3061 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3181 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3241 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3301 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3361 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4081 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5401 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     5461 catagttaag ccagtatctg ctccctgctt g
//
