LOCUS       sequence-384905-        6140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clathrin labelled with near-infrared fluorescent protein 
            miRFP718nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6140)
  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 6140)
  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: pClathrin-miRFP718nano
FEATURES             Location/Qualifiers
     source          1..6140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      179..314
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     241..260
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        376..423
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             442..816
                     /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    946..1067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(983..1002)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1037..1056
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1116..1132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1116..1132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1129..1151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1140..1156
                     /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(1164..1170,1171..1188,1189..1194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1209..1230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1347..1364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1518..2106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1598..1617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2277..3068,3069..3137))
                     /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     2900..2919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3138..3242)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3317..3336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3508..3887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3888..4091
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4042..4062
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4136..4155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4136..4154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5336..5365
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5381..5398
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5421..5438)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5427..5651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5697..6125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5784..5803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5994..6015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(6134..6140,1..14))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6139..6140,1..328)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
       61 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
      121 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
      181 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
      241 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
      301 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
      361 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
      421 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
      481 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
      541 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
      601 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
      661 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
      721 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
      781 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
      841 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
      901 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
      961 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1021 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     1081 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     1141 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1201 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1261 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1321 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1381 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1441 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1501 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1561 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1621 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1681 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1741 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1801 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1861 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1921 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1981 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2041 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2101 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2161 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2221 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2281 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2341 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2401 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2461 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2521 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2581 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2641 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2701 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2761 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2821 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2881 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2941 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3001 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3061 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3121 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3181 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3241 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     3301 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     3361 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     3421 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     3481 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     3541 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     3601 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     3661 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     3721 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     3781 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     3841 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     3901 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     3961 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     4021 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     4081 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     4141 cgactcacta tagggagacc caagctggct agttaagctt ggtaccatgg caaacctgga
     4201 caagatgctg aacaccatcg tgaccgaggt gcgccagttc ctgcaagtgg acagactctg
     4261 cgtgttcaag ttcgaggaag attactccgg caacatcatc tacgaagccg tggacgacgg
     4321 atggttgagc atcctgaaga cccacgtgcg ggactgctac ttcatggaaa ccagaggcga
     4381 ggaatacctg cacggcagat accaggccat cgccgacatc caccaggcca atctggcgga
     4441 gagctataga gacttcctga cccagtacca ggtgcgggcc attgtggctg tccccatcct
     4501 gaagggcaag aagctgtggg gcttgttcag cgcccaccaa ctggccgccc ctcggagctg
     4561 gcaggcctgg gaaatcgagt tcctgaaaca gcaagccgtg gtgatgggca tcgccatcca
     4621 gcagagcgga ggtggtggat caggaggagg tggttctgga ggtggtggat caggaggaag
     4681 atctatggct gatgactttg gcttcttctc gtcgtcggag agtggtgccc cggaggcggc
     4741 ggaggaggac ccggcggccg ccttcctggc ccagcaggag agcgagattg caggcataga
     4801 gaacgacgag ggcttcgggg cacctgccgg cagccatgcg gcccccgcac agccgggccc
     4861 cacgagtggg gctggttctg aggacatggg gaccacagtc aatggagatg tgtttcagga
     4921 ggccaacggt cctgctgatg gctacgcagc cattgcccag gctgacaggc tgacccagga
     4981 gcctgagagc atccgcaagt ggcgagagga gcagaggaaa cggctgcaag agctggatgc
     5041 tgcatctaag gtcacggaac aggaatggcg ggagaaggcc aagaaggacc tggaggagtg
     5101 gaaccagcgc cagagtgaac aagtagagaa gaacaagatc aacaaccggg catccgagga
     5161 ggctttcgtg aaggaatcca aggaggagac cccaggcaca gagtgggaga aggtggccca
     5221 gctatgtgac ttcaacccca agagcagcaa gcagtgcaaa gatgtgtccc gcctgcgctc
     5281 ggtgctcatg tccctgaagc agacgccact gtcccgctag tctagagggc ccttcgaaca
     5341 aaaactcatc tcagaagagg atctgaatat gcataccggt catcatcacc atcaccattg
     5401 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     5461 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     5521 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     5581 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     5641 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5821 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5881 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     6001 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     6061 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     6121 gaattaattc tgtggaatgt
//
