LOCUS       sequence-271522-        5553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clathrin labeling with a near-infrared fluorescent protein emiRFP703
            (enhanced miRFP703).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5553)
  AUTHORS   Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti 
            F, Shemetov AA, Testa I, Verkhusha VV
  TITLE     A set of monomeric near-infrared fluorescent proteins for multicolor
            imaging across scales.
  JOURNAL   Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
  PUBMED    31932632
REFERENCE   2  (bases 1 to 5553)
  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 
            Commun."; date: "2020-01-13"; volume: "11(1):239. doi"; pages: " 
            10.1038/s41467-019-13897-6"
COMMENT     Sequence Label: pemiRFP703-clathrin
FEATURES             Location/Qualifiers
     source          1..5553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        145..448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        449..652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     603..623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2497..2618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2534..2553)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2588..2607
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2625..3080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2762..2783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2974..2993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3107..3211
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3236..3256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3280..3490
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3302..3498
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3349..3484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3533..4327
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMHDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3587..3606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4197..4216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4515..4534)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4559..4606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4935..5523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5424..5443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
       61 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
      121 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      181 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      481 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      541 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      601 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      661 gaaccgtcag atccgctagc caccatggcg gaaggatccg tcgccaggca gcctgacctc
      721 ttgacctgcg aacatgaaga gatccacctc gccggctcga tccagccgca tggcgcgctt
      781 ctggtcgtca gcgaacatga tcatcgcgtc atccaggcca gcgccaacgc cgcggaattt
      841 ctgaatctcg gaagcgtact cggcgttccg ctcgccgaga tcgacggcga tctgttgatc
      901 aagatcctgc cgcatctcga tcccaccgcc gaaggcatgc cggtcgcggt gcgctgccgg
      961 atcggcaatc cctctacgga gtactgcggt ctgatgcatc ggcctccgga aggcgggctg
     1021 atcatcgaac tcgaacgtgc cggcccgtcg atcgatctgt caggcacgct ggcgccggcg
     1081 ctggagcgga tccgcacggc gggttcactg cgcgcgctgt gcgatgacac cgtgctgctg
     1141 tttcagcagt gcaccggcta cgaccgggtg atggtgtatc gtttcgatga gcaaggccac
     1201 ggcctggtat tctccgagtg ccatgtgcct gggctcgaat cctatttcgg caaccgctat
     1261 ccgtcgtcgc tggtcccgca gatggcgcgg cagctgtacg tgcggcagcg cgtccgcgtg
     1321 ctggtcgacg tcacctatca gccggtgccg ctggagccgc ggctgtcgcc gctgaccggg
     1381 cgcgatctcg acatgtcggg ctgcttcctg cgctcgatgt cgccgatcca tctgcagttc
     1441 ctgaaggaca tgggcgtgcg cgccaccctg gcggtgtcgc tggtggtcgg cggcaagctg
     1501 tggggcctgg ttgtctgtca ccattatctg ccgcgcttca tccgtttcga gctgcgggcg
     1561 atctgcaaac ggctcgccga aaggatcgcg acgcggatca ccgcgcttga gagcggaggt
     1621 ggtggatcag gtggaggagg ttctggaggt ggtggatcag gaggaggtgg ttctggaggt
     1681 ggtggatcag gaggaagatc tcgagctcaa gcttcgaatt ctgcagtcga catggctgat
     1741 gactttggct tcttctcgtc gtcggagagt ggtgccccgg aggcggcgga ggaggacccg
     1801 gcggccgcct tcctggccca gcaggagagc gagattgcag gcatagagaa cgacgagggc
     1861 ttcggggcac ctgccggcag ccatgcggcc cccgcacagc cgggccccac gagtggggct
     1921 ggttctgagg acatggggac cacagtcaat ggagatgtgt ttcaggaggc caacggtcct
     1981 gctgatggct acgcagccat tgcccaggct gacaggctga cccaggagcc tgagagcatc
     2041 cgcaagtggc gagaggagca gaggaaacgg ctgcaagagc tggatgctgc atctaaggtc
     2101 acggaacagg aatggcggga gaaggccaag aaggacctgg aggagtggaa ccagcgccag
     2161 agtgaacaag tagagaagaa caagatcaac aaccgggcat ccgaggaggc tttcgtgaag
     2221 gaatccaagg aggagacccc aggcacagag tgggagaagg tggcccagct atgtgacttc
     2281 aaccccaaga gcagcaagca gtgcaaagat gtgtcccgcc tgcgctcggt gctcatgtcc
     2341 ctgaagcaga cgccactgtc ccgctaggga tccaccggat ctagataact gatcataatc
     2401 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2461 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2521 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2581 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3061 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3181 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3361 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3421 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3481 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3541 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3601 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3661 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     3721 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3781 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3841 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3901 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3961 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4021 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcacg acggcgagga
     4081 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4141 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4201 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4261 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4321 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4381 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4441 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4501 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4561 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4621 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4681 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4741 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     4801 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4861 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4921 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5041 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5521 aaaaacgcca gcaacgcggc ctttttacgg ttc
//
