LOCUS       sequence-349255-        6814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Espresses Stonin2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6814)
  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 6814)
  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: GFP-STONIN2
FEATURES             Location/Qualifiers
     source          1..6814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             102..896
                     /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             102..896
                     /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(156..175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     766..785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1084..1103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1128..1175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1504..2092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1993..2012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2267..2570
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2571..2774
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2725..2745
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2813..2822
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2819..2821,2822..2824,2825..3535)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2819..2821,2822..2824,2825..3535)
                     /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(2864..2885)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3125..3144)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3472..3493
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5808..5929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5845..5864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5899..5918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5936..6391)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6073..6094)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6285..6304
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6418..6522
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6524..6814,1..67)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6524..6715
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6547..6567)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(6732..6814,1..53)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
       61 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
      121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
      301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
      661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
      721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
      781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
      841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
      901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
      961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     1021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     1081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     1141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     1201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     1261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     1321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     1381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     1801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     1861 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     1921 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     1981 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2041 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2101 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2161 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     2221 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2281 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2341 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2401 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2461 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2521 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2581 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2641 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2701 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2761 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat
     2821 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2881 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2941 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3001 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3061 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3121 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3181 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3241 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3301 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3361 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3421 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3481 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     3541 actcagatct cgagctcaag cttcgatgga ggccatcagc tggcaggcca gcagtccagc
     3601 catgaatggg caccctgccc ctccagtgac ctctgctcgt tttcccagct gggtcacctt
     3661 tgatgacaat gaagtcagct gccctttacc accagtcacc tctcctctga agccaaatac
     3721 accaccaagt gcatctgtga tcccagatgt accttataat tcaatgggat catttaagaa
     3781 gagggaccgt cccaagagca ctttgatgaa cttctccaaa gttcagaagc tggatatttc
     3841 ctctttaaac cgcacgcctt ctgtaactga ggctccacct tggagggcaa ccaacccttt
     3901 cctgaatgag actctgcagg atgtacagcc ctcccctatc aaccctttca gtgctttctt
     3961 tgaggagcag gagaggcgct cccaaaacag ttccatttcc agtaccacgg gcaaaagcca
     4021 aagagattcc ctcattgtca tctaccagga tgccatcagt tttgatgatt caagcaaaac
     4081 ccagtcacac tctgatgctg ttgaaaaact caaacaactc caaattgatg accctgatca
     4141 ctttggcagt gcaactctac ctgatgatga cccagtagcc tggattgaac tagatgctca
     4201 cccgcctgga tcagcacggt cccagcctcg tgacgggtgg ccaatgatgt tgaggatccc
     4261 tgagaagaaa aacatcatgt cctccaggca ctggggaccg atcttcgtca aactgacaga
     4321 cactggttac ctgcagctgt attatgagca gggcctagaa aaaccattcc gtgagttcaa
     4381 gctggagatc tgtcatgaga tttcagaacc ccggcttcaa aactatgatg agaatggcag
     4441 aatccacagc ttgcggatag accgtgtcac ctataaagag aagaagaaat accagcccaa
     4501 acctgctgtg gcccacacag cagagaggga acaggtgatt aagctgggca ccaccaatta
     4561 cgatgacttc ctgagtttca tccatgcagt tcaggaccgt ctcatggatc tgccagtgtt
     4621 gtcaatggac ttgagcacag ttggcctcaa ctaccttgaa gaggagatta cagtggatgt
     4681 cagagatgaa ttctctggca ttgtgagcaa aggagacaac cagattctcc agcaccatgt
     4741 cttgacacgg atccacatcc tgagtttcct gtctgggctc gcagagtgcc gcctgggcct
     4801 caatgacatc ctcgtcaaag ggaatgaaat agttttgagg caggacatca tgcccaccac
     4861 caccacaaag tggatcaagc tccatgagtg ccgtttccat gggtgtgtgg atgaggatgt
     4921 tttccacaac tcacgggtca ttctgttcaa ccctttggat gcgtgccggt ttgagctaat
     4981 gcggttcagg acagtgtttg ctgagaagac cttgcctttc acactcagga cggccacaag
     5041 tgtcaatggg gcagaggtgg aggtgcagag ctggctgagg atgtcaactg gcttctccgc
     5101 caatcgtgac cccctcactc aggttccctg tgagaatgtg atgatccgtt accctgtgcc
     5161 cagtgagtgg gtgaaaaact tccgcaggga aagtgtcctg ggggaaaagt ctttgaaagc
     5221 caaagtgaac cggggggcaa gttttggctc cactagtgtt tctggctctg agcctgtcat
     5281 gagagtaact ctgggaactg ccaagtacga gcatgccttc aactccattg tgtggaggat
     5341 aaaccgactg ccggacaaaa actcagcttc cggtcaccca cactgtttct tttgccacct
     5401 tgaactcggc tctgaccggg aagtgccttc cagatttgcc aatcacgtga atgtcgagtt
     5461 cagcatgccc acaacttctg cctccaaagc cagcgtgaga tctatctctg tagaagacaa
     5521 gactgacgtc aggaagtggg tcaattattc tgcacactac agctaccagg tagccttggg
     5581 aagcatctgg ctaatgcttc caactccatt tgttcacccc actaccctcc ccctcttatt
     5641 ccttttagcc atgcttacta tgttcgcctg gtaacccggg atccaccgga tctagataac
     5701 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     5941 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6001 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6061 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6121 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6181 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6241 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6301 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6361 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     6421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6481 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     6541 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6601 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     6661 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     6721 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6781 gactaatttt ttttatttat gcagaggccg aggc
//
