LOCUS       sequence-384914-        4846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter of inflamation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  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 4846)
  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: miRFP718nano-based inflammation reporter
FEATURES             Location/Qualifiers
     source          1..4846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        21..212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     169..189
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    406..527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(443..462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     497..516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(579..698)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     regulatory      1145..1154
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1181..1212)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    1328..1419
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(1360..1379)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    1433..1481
                     /note="synthetic polyadenylation signal"
     CDS             join(1586..1654,1655..2446)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2646..3234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3135..3154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3388..3405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3561..3609
                     /note="synthetic polyadenylation signal"
     CDS             complement(3633..4670)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3646..3654
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(join(4701..4846,1..212))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4715..4846,1..4)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4769..4788)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
       61 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
      121 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
      181 acacattcca cagctggttc tttccgcctc agaaggtagc taaccaagtt cctctttcag
      241 aggttatttc aggccatggt gctgcgcaga tcagcggaag agcgcccaat acgcaaacgg
      301 atccttatcg attttaccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
      361 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
      421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
      481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg
      541 aagcggccgg ccgccccgac tctagagtcg cggccttaga cgttgatgcg agctgaagca
      601 caagcggctg ggtgccggtc cataccgctt tcttgtgcgc aggacatagg caatgtaccg
      661 gcggcttgct cttcaacctc aggcggaaag ccgtgagaat tcagctagct ctgctggatg
      721 gcgatgccca tcaccacggc ttgctgtttc aggaactcga tttcccaggc ctgccagctc
      781 cgaggggcgg ccagttggtg ggcgctgaac aagccccaca gcttcttgcc cttcaggatg
      841 gggacagcca caatggcccg cacctggtac tgggtcagga agtctctata gctctccgcc
      901 agattggcct ggtggatgtc ggcgatggcc tggtatctgc cgtgcaggta ttcctcgcct
      961 ctggtttcca tgaagtagca gtcccgcacg tgggtcttca ggatgctcaa ccatccgtcg
     1021 tccacggctt cgtagatgat gttgccggag taatcttcct cgaacttgaa cacgcagagt
     1081 ctgtccactt gcaggaactg gcgcacctcg gtcacgatgg tgttcagcat cttgtccagg
     1141 tttgccatgg tggctttacc aacagtaccg gattgccaag ctggaagtcg agcttccatt
     1201 atataccctc tagtgtctaa gcttggccgc cgaggccaga tctggaaatt cccggaaagt
     1261 ccccggaaat tcccggaaag tccccggaaa ttcccgctag cgagctcagg taccggccag
     1321 ttaggccaga gaaatgttct ggcacctgca cttgcactgg ggacagccta ttttgctagt
     1381 ttgttttgtt tcgttttgtt ttgatggaga gcgtatgtta gtactatcga ttcacacaaa
     1441 aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggcctgt ccaatacctc
     1501 ccgtacctta atattactta cttatccttg agagacgtac tagtaaccct gataaatgct
     1561 tcaataatat tgaaaaagga cgagtatgag catccaacat tttcgtgtcg cactcattcc
     1621 cttctttgcg gcattttgct tgcctgtttt tgcacacccc gaaacgctgg tgaaagtaaa
     1681 agatgctgaa gatcaactgg gtgcaagagt gggctatatc gaactggatc tcaatagcgg
     1741 caagatcctt gagtctttcc gccccgaaga acgattcccg atgatgagca ctttcaaagt
     1801 actgctatgt ggcgcggtgt tgtcccgtat agacgccggg caagagcagc ttggtcgccg
     1861 tatacactac tcacaaaacg acttggttga gtactcgccg gtcacggaaa agcatcttac
     1921 ggatggcatg acggtaagag aattgtgtag tgctgccatt accatgagcg acaacaccgc
     1981 ggccaactta cttctgacaa cgatcggagg ccctaaggag ctgactgcat ttcttcataa
     2041 tatgggtgat catgtgaccc ggcttgaccg ctgggaacca gagttgaacg aagccatacc
     2101 gaacgacgag cgtgatacca cgatgccagt agcaatggcc acaactcttc ggaaactact
     2161 cactggcgaa cttcttactc tagcatcacg acagcagctc atagactgga tggaggcgga
     2221 caaagtagca ggaccacttc ttcgctcggc cctccctgct ggctggttca ttgctgacaa
     2281 atcgggggcc ggtgaacgcg gctctcgcgg catcattgct gcgctggggc ctgatggtaa
     2341 gccctcacga atcgtagtga tctacacgac ggggagtcag gccactatgg acgaacgaaa
     2401 taggcagatc gctgagatcg gtgcctcact gatcaagcac tggtaaccac tgcagtggtt
     2461 tagcatttgc ggccgctgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2521 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2581 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2641 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2701 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2761 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2821 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2881 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2941 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3001 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3061 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3121 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3181 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3241 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3301 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3361 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3421 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     3481 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3541 tcgacagcat ctagttcgaa cacacaaaaa accaacacac agatgtaatg aaaataaaga
     3601 tattttattg cggccatgac cggttctaga gtttaaactc gacctacctc cttagcgcgc
     3661 ggtcgtgtac tgggccggcg gttgccgctg tcggccagca cctcgacgca gccgtcggtc
     3721 catacggctg cgctccggcg agcgatttgt gtgcgaccga cggtgccggc cccgctgcgg
     3781 acgatggcat cgcagcggcc ttgagcccag gcagcatcgt cgaagttgcc gtccacgagg
     3841 ctctggtaga gctgatccag gccgatgcgc agcatgtagg ctcgcagacg agggctgccg
     3901 gccagctcgg gatgccggcg ctcgaagtag cgagtctgct gctccatgca agccagccag
     3961 ggccgccaga agaagatgtt ggccacctcg tactgactgt ccccgaacat agcttcggac
     4021 cagtcgatta cggcggtgat gcggccgttg tcggtcagga cgttgttgct gccgaagtcg
     4081 gcatggacga ggtggcgcac ctcgggacag tcttcggccc acagcatcag ttcgtccagg
     4141 gcttgagcta cgctggcgga cacggtgtcg tccatcacgg tctgccagtg gtagacatgg
     4201 ggatcagcaa tggcgcaaat gaaatcccgc caggttgtgt actggccgat gccttggggc
     4261 ccgaatgggc cgaagccgct ggtttgactg aggtcggcgg cggcaatagc atccatagct
     4321 tcggcgacag gctgtaacac agcaggcagc tctgtttcgg ggaggtcttg gagagtaacg
     4381 ccttgggcgc gtctactgat gcagtatgtc aggctctcgc tgaactcgcc gatgtccaac
     4441 acttcgggga tgggtagtgc agcgctggcg aagtggcggt acacgtagcg gtctttgtag
     4501 aagccatcag cgcagctatt cacccgcagt acatagccgc gtccgccgac atcgaagctg
     4561 aaggctcggc tctcttcgcc ctccgacaac tgcatcaggt cgctcacact gtcgaacttc
     4621 tcgatgagaa atttttcaac gctggtagcg gtgagttcgg gcttcttcat ggtggcgcta
     4681 gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     4741 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
     4801 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcg
//
