LOCUS       sequence-218573-        5313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Clathrin Vesicles, Excitation: 503 / 569, Emission: 
            514 / 581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 5313)
  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"
COMMENT     Sequence Label: mGeos2-VEL-Clathrin-15
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             671..1348
                     /codon_start=1
                     /product="truly monomeric reversibly photoswitchable 
                     fluorescent protein, suitable for nonlinear structured 
                     illumination microscopy (Zhang et al., 2016)"
                     /label=Skylan-NS
                     /note="mEos3.1 derivative that eliminates the green-to-red 
                     conversion"
                     /translation="MSAIKPDMKIKLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFLYGNRVFAKYPDNIQDYFKQSFPKGYSWERSLTFEDGGICNAR
                     NDITMEGDTFYNKVRFYGTNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDVEMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGAHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     polyA_signal    2159..2280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2196..2215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2250..2269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2287..2742)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2424..2445)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2636..2655
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2769..2873
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2875..3232
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2875..3066
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2898..2918)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3083..3218
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3267..4061
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3267..4061
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3321..3340)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3931..3950
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4249..4268)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4293..4340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4669..5257
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5158..5177
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atgagtgcga ttaagccaga catgaagatc aaactccgta tggaaggcaa
      721 cgtaaacggg caccactttg tgatcgacgg agatggtaca ggcaagcctt ttgagggaaa
      781 acagagtatg gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct
      841 gaccactgca ttcctgtacg gcaacagggt attcgccaaa tatccagaca acatacaaga
      901 ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa cgaagcttga ctttcgaaga
      961 cgggggcatt tgcaacgcca gaaacgacat aacaatggaa ggggacactt tctataataa
     1021 agttcgattt tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct
     1081 gaaatgggag ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatgt
     1141 agagatggct ttgttgcttg aaggaaatgc ccattaccga tgtgacttca gaactactta
     1201 caaagctaag gagaagggtg tcaagttacc aggcgcccac tttgtggacc actgcattga
     1261 gattttaagc catgacaaag attacaacaa ggttaagctg tatgagcatg ctgttgctca
     1321 ttctggattg cctgacaatg ccagacgatc cggactcaga tctcgagctc aagcttcgaa
     1381 ttctgcagtc gacatggctg atgactttgg cttcttctcg tcgtcggaga gtggtgcccc
     1441 ggaggcggcg gaggaggacc cggcggccgc cttcctggcc cagcaggaga gcgagattgc
     1501 aggcatagag aacgacgagg gcttcggggc acctgccggc agccatgcgg cccccgcaca
     1561 gccgggcccc acgagtgggg ctggttctga ggacatgggg accacagtca atggagatgt
     1621 gtttcaggag gccaacggtc ctgctgatgg ctacgcagcc attgcccagg ctgacaggct
     1681 gacccaggag cctgagagca tccgcaagtg gcgagaggag cagaggaaac ggctgcaaga
     1741 gctggatgct gcatctaagg tcacggaaca ggaatggcgg gagaaggcca agaaggacct
     1801 ggaggagtgg aaccagcgcc agagtgaaca agtagagaag aacaagatca acaaccgggc
     1861 atccgaggag gctttcgtga aggaatccaa ggaggagacc ccaggcacag agtgggagaa
     1921 ggtggcccag ctatgtgact tcaaccccaa gagcagcaag cagtgcaaag atgtgtcccg
     1981 cctgcgctcg gtgctcatgt ccctgaagca gacgccactg tcccgctagg gatccaccgg
     2041 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2101 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2161 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2221 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2281 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2341 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2401 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2461 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2521 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2581 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2641 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2701 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2881 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3061 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3121 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3181 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     3241 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3301 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3361 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3421 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     3481 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3541 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3601 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3661 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3721 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3781 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3841 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3901 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3961 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4021 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4081 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4201 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     4261 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     4321 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4381 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4441 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     4501 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5281 acggttcctg gccttttgct ggccttttgc tca
//
