LOCUS       sequence-368437-        7369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the fusion protein FKBP-AP180-GFP. Uses rat AP180 
            residues 328-896..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7369)
  AUTHORS   Kuey C, Sittewelle M, Larocque G, Hernandez-Gonzalez M, Royle SJ
  TITLE     Recruitment of clathrin to intracellular membranes is sufficient for
            vesicle formation.
  JOURNAL   Elife. 2022 Jul 19;11. pii: 78929. doi: 10.7554/eLife.78929.
  PUBMED    35852853
REFERENCE   2  (bases 1 to 7369)
  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: "Elife."; 
            date: "2022-07-19"; volume: "11. pii: 78929. doi"; pages: " 
            10.7554/eLife.78929"
COMMENT     Sequence Label: FKBP-AP180-GFP
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(666..668,669..989)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     regulatory      3309..3318
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3315..3317,3318..3320,3321..4034)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3315..3317,3318..3320,3321..4031)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3360..3381)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3621..3640)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3968..3989
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4157..4278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4248..4267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4285..4740)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4422..4443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4634..4653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4767..4871
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4873..5230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4873..5064
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4896..4916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5081..5216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5265..6059
                     /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             5265..6059
                     /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(5319..5338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5929..5948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6247..6266)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6291..6338
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6667..7255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7156..7175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagag atcgatgtga atgggggccc tgaacccgcc aagaacctgt
      661 acaccatggg agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc
      721 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt
      781 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
      841 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
      901 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
      961 tcttcgatgt ggagcttcta aaactggaac ggtaccgcgg gcccgggatc cccgtcgaca
     1021 tctttgcaac agcatccgcg gctgccccag tcagctctgc taagccatca agcgatctcc
     1081 ttgatcttca gccagacttc tccggagcag ctgcgggggc agcagcgcct gtaccaccac
     1141 ctactggggg agccactgcg tggggagacc ttttgggaga ggattccttg gctgcacttt
     1201 cctctgttcc ctctgaagca ccgatttctg acccatttgc accagaacct tcccctccta
     1261 ctacaaccac tgaacctgct tcagcttctg cctcagccac cacagctgtg acagctgcca
     1321 ctacagaagt ggatctcttt ggagatgcct ttgcagcttc tcctggggag gcccctgcag
     1381 catccgaagg ggccacggca ccagctaccc cggccccggt ggctgcagct cttgatgcat
     1441 gctcaggaaa tgaccctttt gccccatctg aaggtagcgc agaggctgca cctgagctcg
     1501 acctctttgc aatgaagcca cctgagacca gcgctcctgt agttacccct acagctagca
     1561 cagcccctcc agttcccgca actgctcctt ctcctgctcc cacggctgtg gcagccactg
     1621 ctgccgccac caccaccacc accgccgctg cagctacgac cactgccacc acctctgctg
     1681 ctgctgctgc caccgctgct gctcctcctg ctctagatat ctttggtgat ttgtttgatt
     1741 ctgctcctga agttgctgcg gcatctaagc cggacgtggc tcctagcata gacctgtttg
     1801 gcacagacgc tttctcctcc ccaccacgag gggcctctcc ggtgcctgag agttctctca
     1861 ctgctgacct cttatccgtg gacgcgtttg cagcgccgtc gcctgcatcc actgcctctc
     1921 ctgcaaaggc ggagtcctcg ggtgtcatag acctttttgg ggatgcgttt ggaagtagtg
     1981 cttctgaaac ccagccagca ccacaggctg tttctagttc atcagcatcg gcagatctac
     2041 tggctggatt tgggggttct ttcatggccc cttctacaac acctgtgact ccagctcaga
     2101 ataacctact gcaacccaat tttgaggcag cttttggaac aacgccttcg acttcaagcc
     2161 cttctcctgc tcccacggct gtggcagcca ctgctgccgc caccaccacc accaccgccg
     2221 ctgcagctac gaccactgcc accacctctg ctgctgctgc tgccaccgct gctgctcctc
     2281 ctgctctaga tatctttggt gatttgtttg attctgctcc tgaagttgct gcggcatcta
     2341 agccggacgt ggctcctagc atagacctgt ttggcacaga cgctttctcc tccccaccac
     2401 gaggggcctc tccggtgcct gagagttctc tcactgctga cctcttatcc gtggacgcgt
     2461 ttgcagcgcc gtcgcctgca tccactgcct ctcctgcaaa ggcggagtcc tcgggtgtca
     2521 tagacctttt tggggatgcg tttggaagta gtgcttctga aacccagcca gcaccacagg
     2581 ctgtttctag ttcatcagca tcggcagatc tactggctgg atttgggggt tctttcatgg
     2641 ccccttctac aacacctgtg actccagctc agaataacct actgcaaccc aattttgagg
     2701 cagcttttgg aacaacgcct tcgacttcaa gcagcagctc cttcgaccca tcaggcgatc
     2761 ttctgatgcc aaccatggca ccgtccgggc agcctgcccc tgtctcaatg gtcccaccca
     2821 gtcctgcaat gtcagccagc aaaggccttg gaagtgacct tgactcgtct ctggccagct
     2881 tagtaggcaa tcttgggatt tctggtacca catcaaaaaa gggagatctc cagtggaatg
     2941 ctggggagaa gaagttgacc ggtggagcca actggcaacc aaaagtcact ccggccacat
     3001 ggtcagcagg cgttccaccg caaggcactg ttcctccaac cagctcagtc cctccgggtg
     3061 ccggcgcccc gtctgttggg caacctggag caggatatgg aatgcctcct gcggggacag
     3121 gcatgaccat gatgcctcag cagccagtca tgtttgcaca gcccatgatg agaccaccct
     3181 ttggagctgc agctgtgccc ggcacacagc tttctccaag ccctacacct gccactcaga
     3241 gtcccaagaa acctccagcc aaggacccat tagcggatct taacatcaag gatttcttgc
     3301 tcccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3361 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     3421 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     3481 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3541 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3601 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3661 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3721 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     3781 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3841 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3901 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     3961 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     4021 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     4081 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4141 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4201 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4261 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4321 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4381 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4441 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4501 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4561 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4621 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4681 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4741 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4801 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4861 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4921 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4981 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5041 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5101 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5161 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5221 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5281 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5341 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5401 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5461 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5521 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5581 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5641 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5701 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5761 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5821 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5881 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     5941 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6001 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6061 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6121 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6181 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     6241 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6301 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6361 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6421 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6481 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6541 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6601 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6781 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
