LOCUS       sequence-337775-        5788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Endophilin A1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5788)
  AUTHORS   Feng Z, Yu CH
  TITLE     PI(3,4)P2-mediated membrane tubulation promotes integrin trafficking
            and invasive cell migration.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 May 11;118(19). pii: 2017645118. doi:
            10.1073/pnas.2017645118.
  PUBMED    33947811
REFERENCE   2  (bases 1 to 5788)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-05-11"; volume: "118(19). pii: 
            2017645118. doi"; pages: " 10.1073/pnas.2017645118"
COMMENT     Sequence Label: EndophilinA1-mRuby2
FEATURES             Location/Qualifiers
     source          1..5788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1742..1751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1748..2458
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     polyA_signal    2583..2704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2620..2639)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2674..2693
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2711..3166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2848..2869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3060..3079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3193..3297
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3299..3656
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3299..3490
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3322..3342)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3507..3642
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3691..4485
                     /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             3691..4485
                     /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(3745..3764)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4355..4374
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4673..4692)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4717..4764
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5093..5681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5582..5601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttatgtcg gtggccggcc tcaagaagca
      661 gttccataaa gccactcaga aagtgagtga gaaggttgga ggagctgaag gaaccaagct
      721 agatgatgac ttcaaagaga tggaaaggaa agtggatgtc accagcaggg ctgtgatgga
      781 aataatgact aaaacaattg aataccttca acccaatcca gcttccagag ctaagctcag
      841 catgatcaac accatgtcaa aaatccgtgg ccaggagaag gggccaggct atcctcaggc
      901 agaggcgctg ctggcagagg ccatgctcaa atttggaaga gagcttggag atgattgcaa
      961 ctttggccca gcacttggtg aggtcgggga ggccatgcgg gaactgtcgg aggtcaaaga
     1021 ctctttggac atagaagtga agcagaactt cattgaccct cttcagaatc ttcatgacaa
     1081 agatcttagg gaaattcaac atcatctaaa gaagttggag ggtcgacgcc tggattttga
     1141 ttataagaag aaacgacaag gcaagattcc ggatgaagag cttcgtcaag ctctagagaa
     1201 atttgatgag tctaaggaaa ttgctgagtc aagcatgttc aatctcttgg agatggatat
     1261 tgaacaagtg agccagctct ctgcacttgt gcaagctcag ctggagtacc acaagcaggc
     1321 agtccagatc ctgcagcaag tcacggtcag actggaagaa agaataagac aggcttcatc
     1381 tcagcctaga agggaatatc aacctaaacc acgaatgagc ctggagtttc caactggaga
     1441 cagtactcag cccaatgggg gtctctccca cacaggcact cccaaacctt caggtgtcca
     1501 aatggatcag ccctgctgcc gagctctgta cgactttgaa cctgaaaatg aaggggagtt
     1561 gggatttaaa gagggcgata tcatcacact cactaaccaa attgatgaga actggtatga
     1621 ggggatgctg catggccatt caggcttctt ccccatcaat tatgtggaaa ttctggttgc
     1681 cctgccccat aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt
     1741 cgccaccatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
     1801 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaatcc
     1861 gtacatggga actcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     1921 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccgaa
     1981 aggcattcct gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac
     2041 gagatacgaa gatggtggag tcgtcaccgt catgcaggac accagccttg aggatggctg
     2101 tctcgtttac cacgtccaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     2161 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     2221 gaggggatac actcatatgg cactgaaagt tgatggtggt ggccatctgt cttgctcttt
     2281 cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtatccatgc
     2341 cgttgatcac cgcctggaaa ggttagagga aagtgacaat gaaatgttcg tagtacaacg
     2401 cgaacacgca gttgccaagt tcgccgggct tggtggtggg atggacgagc tgtacaagta
     2461 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2521 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2581 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2641 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2701 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2761 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2821 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2881 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2941 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3001 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3061 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3121 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3181 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3241 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3301 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3481 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3541 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3601 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3661 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3721 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3781 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3841 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3901 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3961 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4021 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4081 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4141 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4201 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4261 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4321 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4381 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4441 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4501 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4561 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4621 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4681 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4741 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4801 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4861 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4921 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4981 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5041 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5101 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5161 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5221 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5281 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5341 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5401 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5461 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5521 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5641 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5701 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5761 ctgattctgt ggataaccgt attaccgc
//
