LOCUS       sequence-410755-        6170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clathrin labelled with near-infrared fluorescent protein 
            miRFP704nano.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6170)
  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 6170)
  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: pClathrin-miRFP704nano
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     90..109
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        225..272
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             291..665
                     /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    795..916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(832..851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     886..905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(965..981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(965..981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(978..1000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    989..1005
                     /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(1013..1019,1020..1037,1038..1043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1058..1079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1196..1213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1367..1955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1447..1466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2126..2917,2918..2986))
                     /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     2749..2768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2987..3091)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3166..3185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3357..3736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3737..3940
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3891..3911
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3985..4004
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3985..4003
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5215..5244
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5260..5277
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5300..5317)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5306..5530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5576..6004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5663..5682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5873..5894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6013..6033)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6018..6170,1..177)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
       61 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
      121 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
      181 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
      241 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
      301 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
      361 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
      421 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
      481 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
      541 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
      601 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
      661 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
      721 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
      781 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
      841 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      901 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
      961 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1021 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1081 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1141 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1201 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1801 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2041 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2101 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3001 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3061 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     3361 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3421 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3481 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3541 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3601 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3661 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3721 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3781 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3841 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3901 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     3961 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     4021 gttaagcttg gtaccatggc aaacctggac aagatgctga acaccatcgt gaccgaggtg
     4081 cgccagttcc tgcaagtgga cagagtgtgc gtgttccagt tcgaggaaga ttactccggc
     4141 agcgtcgtcg tggaagccgt ggacgacaga tggaacagca tcctgaagac ccaggtgcgg
     4201 gactgctact tcatggaaac cagaggtgag gaatacttgc acggcagata ccaggccatc
     4261 gccgacatct accaggccaa tctgctggag agctacagag acctgctggg ccagtttcag
     4321 gtgcgggcca ttctggctgt ccccatcatc aagggcaaga agctgtgggg cctgctggtg
     4381 gcccaccaac tggctgcccc tcggagctgg cagacctggg aaatcgagtt cctgaaacag
     4441 caagccgtgg tgatgggcat cgccatccag cagagcggag gtggtggatc aggaggaggt
     4501 ggttctggag gtggtggatc aggaggaggt ggttctggag gtggtggatc aggaggaaga
     4561 tctatggctg atgactttgg cttcttctcg tcgtcggaga gtggtgcccc ggaggcggcg
     4621 gaggaggacc cggcggccgc cttcctggcc cagcaggaga gcgagattgc aggcatagag
     4681 aacgacgagg gcttcggggc acctgccggc agccatgcgg cccccgcaca gccgggcccc
     4741 acgagtgggg ctggttctga ggacatgggg accacagtca atggagatgt gtttcaggag
     4801 gccaacggtc ctgctgatgg ctacgcagcc attgcccagg ctgacaggct gacccaggag
     4861 cctgagagca tccgcaagtg gcgagaggag cagaggaaac ggctgcaaga gctggatgct
     4921 gcatctaagg tcacggaaca ggaatggcgg gagaaggcca agaaggacct ggaggagtgg
     4981 aaccagcgcc agagtgaaca agtagagaag aacaagatca acaaccgggc atccgaggag
     5041 gctttcgtga aggaatccaa ggaggagacc ccaggcacag agtgggagaa ggtggcccag
     5101 ctatgtgact tcaaccccaa gagcagcaag cagtgcaaag atgtgtcccg cctgcgctcg
     5161 gtgctcatgt ccctgaagca gacgccactg tcccgctagt ctagagggcc cttcgaacaa
     5221 aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca tcaccattga
     5281 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5341 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5401 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5461 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5521 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     5581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5641 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5761 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5881 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5941 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6001 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6061 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
//
