LOCUS       sequence-348377-        7008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Stonin1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7008)
  AUTHORS   Martina JA, Bonangelino CJ, Aguilar RC, Bonifacino JS
  TITLE     Stonin 2: an adaptor-like protein that interacts with components of 
            the endocytic machinery.
  JOURNAL   J Cell Biol. 2001 May 28;153(5):1111-20. doi: 
            10.1083/jcb.153.5.1111.
  PUBMED    11381094
REFERENCE   2  (bases 1 to 7008)
  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"; doi: "10.1083/jcb.153.5";
            journalName: "J Cell Biol"; date: "2001-05-28- 28"; volume: "153"; 
            issue: "5"; pages: "1111-20"
COMMENT     Sequence Label: pEGFP-HA3-STONIN1
FEATURES             Location/Qualifiers
     source          1..7008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        110..413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     568..588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      656..665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(662..664,665..667,668..1378)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(662..664,665..667,668..1378)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(707..728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(968..987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1315..1336
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1412..1438
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1442..1468
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1475..1501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3845..3966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3882..3901)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3936..3955
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3973..4428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4110..4131)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4322..4341
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4455..4559
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4561..4918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4561..4752
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4584..4604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4769..4904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4953..5747
                     /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             4953..5747
                     /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(5007..5026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5617..5636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5935..5954)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5979..6026
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6355..6943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6844..6863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
       61 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      121 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      181 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      241 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      301 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      361 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      421 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      481 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      541 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      601 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac
      661 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      721 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      781 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      841 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      901 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      961 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1021 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1081 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1141 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1201 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1261 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1321 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     1381 cggccggact cagatctcga gcgccaccat gtacccgtac gatgttcctg actatgcggg
     1441 ctatccctat gacgtcccgg actatgccat gggctaccct tacgacgttc cagattacgc
     1501 tggtaccatg tgctccacaa atccaggcaa atgggtcacc tttgatgatg atcctgctgt
     1561 tcaatcttct caaaagtcaa agaattttcc tctggagaat caaggtgtct gtagaccaaa
     1621 tggactgaag ctgaaccttc ctggcctcag ggaatttccc agtggatctt cctccaccag
     1681 cagcactcct ctctcctccc ccattgtaga tttttatttc agtccaggac ctccaagtaa
     1741 ctctcctctt tctacaccta ccaaagactt cccaggtttt cctggcatcc ccaaagcagg
     1801 gactcatgtg ctttatccta ttccagaatc atcttcagac agcccactcg caatatcagg
     1861 aggagaatct tccttactgc ctaccagacc aacatgttta tcccatgcct tgttacccag
     1921 tgaccactca tgtacacatc caactcccaa agtaggtctt ccagatgaag ttaatcctca
     1981 acaggctgaa agcctaggat tccaaagtga tgatctcccc cagtttcagt attttcgaga
     2041 ggactgtgct ttttcaagtc cattttggaa agatgaaggc agtgattccc atttcaccct
     2101 tgacccacca ggaagcaaaa agatgttctc atcaagaaac aaggagatgc ctattgacca
     2161 aaaaagccta aataagtgtt cactcaacta tatctgtgag aagcttgaac atctccagtc
     2221 agctgagaac caagactcac ttagaagttt gtctatgcac tgtctatgtg ctgaagaaaa
     2281 tgcctcttcc tttgtccccc acacactctt caggagtcag ccaaaatccg gatggtcttt
     2341 catgctgaga attcctgaga agaagaatat gatgtcttcc cggcaatggg gaccaatttt
     2401 tctgaaagtt ttgcctggag gaattttgca gatgtattat gaacagggat tagaaaaacc
     2461 atttaaagag atacagcttg atccatattg taggctttct gaacccaagg ttgagaactt
     2521 cagtgtagca ggaaaaatcc acactgtgaa gattgaacat gtgtcttaca cagaaaaaag
     2581 gaaataccat tctaagacag aagtagttca tgaacctgac atagagcaga tgctgaagtt
     2641 ggggtccaca tcgtaccatg acttccttga ctttctgact actgtggagg aggagctgat
     2701 gaagttgcca gctgtttcaa aaccaaaaaa gaactacgag gagcaagaaa tttccttgga
     2761 aattgtggac aacttttggg gtaaagtcac aaaagaagga aaatttgctg aaagtgctgt
     2821 gataactcaa atttattgcc tctgctttgt gaatgggaac ctggaatgct ttttaacctt
     2881 gaatgacctt gagttgccga agcgagatga atcctattat gagaaggact cagaaaaaaa
     2941 ggggattgat attcttgact accattttca taagtgtgtg aatgtacaag aatttgagca
     3001 atcaagaatc attaagtttg tacctctgga tgcctgccgg tttgagctga tgcgtttcaa
     3061 gactttgtat aatggggata atcttccctt ttccttgaag tctgtagtgg ttgtccaggg
     3121 agcatacgtg gaacttcagg cttttgtcaa catggcctca ttggcgcaga ggtcatccta
     3181 cgctggttcc ttaaggtcct gtgacaatat aaggatacac tttcctgtcc catcgcagtg
     3241 gatcaaggcc ctttggacca tgaacctcca gaggcagaag tctctgaaag ctaaaatgaa
     3301 ccgccgagca tgtctgggga gtttacagga acttgaatct gaacctgtca ttcaagtcac
     3361 tgtggggtca gcaaaatatg agagtgccta ccaggcagtg gtatggaaga tagatcggct
     3421 tccagacaaa aattcaagtc tagatcatcc ccattgtctg tcatacaaat tagagcttgg
     3481 atcagaccaa gaaattccct ctgattggta tccatttgct actgttcagt tttccgtgcc
     3541 tgacacctgt gcctcaagga cagaggtcag gtctctggga gtggagagtg atgtccagcc
     3601 acagaaacat gttcagcagc gagcttgcta caacatccag gttgaaatag aaaaaaagtg
     3661 gattaaaatc gatggagaag acccagataa aattggtgac tgcataactc agtagggatc
     3721 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3781 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3841 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3901 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3961 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4021 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4081 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4141 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4201 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4261 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4321 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4381 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4441 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4501 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4561 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4621 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4741 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4801 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4861 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4921 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4981 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5041 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5101 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5161 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5221 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5281 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5341 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5401 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5461 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5521 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5581 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5641 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5701 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5761 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5821 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5881 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5941 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6001 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6061 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6121 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6181 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6241 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6301 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6361 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6421 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6481 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6541 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6601 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6661 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6721 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6781 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6841 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6901 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6961 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctt
//
