LOCUS       sequence-337974-        7635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses INPP4A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7635)
  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 7635)
  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: EGFP-INPP4A
FEATURES             Location/Qualifiers
     source          1..7635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        60..363
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        364..567
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     518..538
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(612..614,615..617,618..1328)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(612..614,615..617,618..1328)
                     /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(657..678)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(918..937)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1265..1286
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4423..4544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4460..4479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4514..4533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4551..5006)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4688..4709)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4900..4919
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5033..5137
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5139..5496
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5139..5330
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5162..5182)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5347..5482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5531..6325
                     /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             5531..6325
                     /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(5585..5604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6195..6214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6513..6532)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6557..6604
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6933..7521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7422..7441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
      601 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      661 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      721 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      781 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      841 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      901 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      961 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1021 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1081 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1141 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1201 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1261 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1321 tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttctatgaca gcaagagagc
     1381 acagccctcg ccatggtgcc agggcccgtg caatgcagcg ggcttccacc atcgacgtgg
     1441 cggccgacat gctgggcctc tctctggcag gaaatataca agacccagat gagcccattt
     1501 tagaatttag cttagcttgc agtgagctgc atactccatc gctagatcga aagccaaata
     1561 gttttgttgc ggtgagtgtc accacccctc ctcaggcatt ctggacgaag catgcacaga
     1621 cggagatcat tgagggaacc aacaatccta tatttctaag cagtattgcc ttctttcaag
     1681 actctcttat caatcagatg acacaagtca aactctccgt gtatgatgtc aaagatagat
     1741 ctcagggaac aatgtattta ctgggctctg gaacgttcat tgtcaaagat ctgctccagg
     1801 acaggcatca taggttgcat ttaacactaa ggtctgcaga gagtgaccgt gtaggtaaca
     1861 tcaccgtgat tggctggcag atggaggaga agtcagacca acggccccct gtgacccggt
     1921 ctgtggacac tgtcaatggg aggatggttc ttcctgtcga tgagagcttg acggaggcgt
     1981 taggaatccg atccaaatac gcttcattgc gaaaggacac tttgctgaaa tcggtgttcg
     2041 gtggtgccat ctgccgcatg taccggtttc caaccactga tggtaaccat ttgcggatcc
     2101 tggagcagat ggcagagagc gtgctctccc tgcacgtgcc ccggcagttc gtgaagctcc
     2161 tactagagga agatgcagcc agagtgtgtg agctggagga gctgggagag ctgtcccctt
     2221 gctgggagag cctccggcgc caaattgtca cccagtacca gaccatcatc ctcacatacc
     2281 aggagaacct gaccgacctc catcagtaca gagggccctc gtttaaagca agcagtttga
     2341 aagcagataa aaagttagaa tttgttccca caaacttgca catacaaagg atgagagttc
     2401 aagacgatgg aggatcagat cagaactacg acatcgtcac cattggggcg ccagcagcac
     2461 actgccaagg ttttaagtca ggaggtctcc gcaaaaagct gcacaaattt gaagagacca
     2521 agaaacatac atcatctggc tgccagtcca taatctacat accccaggat gttgtcagag
     2581 ccaaggagat catcgcccag atcaacaccc tgaaaaccca agtgagttac tacgcagagc
     2641 ggctgtcaag ggcagccaag gacaggtctg ccactggcct tgagaggaca ctcgccatct
     2701 tggcagacaa gacacggcag ctggtcacgg tctgcgactg caagctcctg gccaactcca
     2761 tccatgggct gaacgctgca cggcctgact acattgcctc caaggcctct cccacttcga
     2821 ctgaggagga gcaggtgatg cttagaaatg accaggacac cctcatggcc cggtggacag
     2881 ggaaaaacag ccgatcttcc ctgcaggtgg actggcacga ggaggagtgg gagaaagtgt
     2941 ggctgaacgt ggacaagagc ctagagtgca tcattcagcg tgtggacaag ctgctgcaga
     3001 aggagcggct gcatggcgag ggctgtgagg atgtcttccc ctgtgcaggc agctgcacca
     3061 gcaagaaagg taacccggac agccacgcct actggatcag accagaagac cccttctgtg
     3121 atgtcccctc ctcaccatgc ccctccacca tgccctccac tgcatgccat cctcatctga
     3181 ccacacattg cagtccccct cctgaagagt ccagcccagg tgaatggagt gaggcccttt
     3241 acccgctgct gaccactctc accgactgcg tggccatgat gagtgacaag gccaagaagg
     3301 ccatggtatt cctgctcatg caggacagcg cgcccaccat agccacctac ctgagcctgc
     3361 agtaccgccg tgacgtggtc ttctgccaga cgctgaccgc cctcatctgc ggcttcatca
     3421 ttaagctgag gaactgcctg catgacgacg gcttcctgcg ccagctctac accatcgggc
     3481 tgctggccca gttcgagagc ctgctgagca cctacgggga ggagctggca atgctggagg
     3541 acatgagcct tgggatcatg gacttgagga acgtgacctt caaagtcact caggccactt
     3601 ccagcgcctc cgcagacatg ctgcccgtca tcacaggaaa tcgcgacgag tttaacgtgc
     3661 gggtccctct gccgggcccg ctgtttgacg ccttgccccg ggagatccag agtggcatgc
     3721 tgctgcgagt gcagcccgtc ctcttcaacg tgggcatcaa tgagcagcag acactggccg
     3781 agaggtttgg cgatacgtct ttacaagaag tcatcaacgt ggagagtttg gtgcggttaa
     3841 attcctactt tgagcagttt aaggaagttt tgcctgagga ttgcctgcct cggtctcgca
     3901 gtcagacgtg cctgccagag ctgctgcggt ttctgggtca gaacgtgcat gcccggaaga
     3961 ataagaacgt cgacattctc tggcaagctg ctgagatctg ccgccgcctt aatggggtcc
     4021 ggttcaccag ctgcaagagc gctaaggacc gtacagccat gtcggtgaca ctggagcagt
     4081 gcctgatcct gcaacacgag catggcatgg ccccgcaggt cttcacccag gccctggagt
     4141 gcatgcgcag tgagggttgt cgaagagaaa atacaatgaa gaatgttgga agtcgcaaat
     4201 atgcatttaa ttccctgcag ctgaaggctt tccccaagca ttacaggcct cccgaaggga
     4261 cttacggaaa agttgaaacg gcccgcgggc ccgggatcca ccggatctag ataactgatc
     4321 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4381 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4561 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4621 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4681 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4741 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4801 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4861 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4921 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4981 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     5041 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5101 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5161 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5221 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5401 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5461 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5521 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5581 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5641 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5701 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5761 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5821 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5881 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5941 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6001 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     6061 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6121 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6181 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6241 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6301 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6361 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6421 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6481 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6541 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6601 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6661 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6721 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7621 attaccgcca tgcat
//
