LOCUS       sequence-9514-ex        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6393)
  AUTHORS   Hammond GR, Sim Y, Lagnado L, Irvine RF
  TITLE     Reversible binding and rapid diffusion of proteins in complex with 
            inositol lipids serves to coordinate free movement with spatial 
            information.
  JOURNAL   J Cell Biol. 2009 Jan 26;184(2):297-308. doi: 10.1083/jcb.200809073.
            Epub 2009 Jan 19.
  PUBMED    19153221
REFERENCE   2  (bases 1 to 6393)
  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.200809073"; journalName: "J Cell Biol"; date: 
            "2009-01-26- 26"; volume: "184"; issue: "2"; pages: "297-308"
COMMENT     Sequence Label: pEGFP-C1-GAP1IP4BPdeltaC2
FEATURES             Location/Qualifiers
     source          1..6393
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3218..3237)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3272..3291
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3309..3764)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3446..3467)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3658..3677
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3791..3895
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3897..4254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3897..4088
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3920..3940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4105..4240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4289..5083
                     /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             4289..5083
                     /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(4343..4362)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4953..4972
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5271..5290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5315..5362
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct cgcctaaagc cagacgacct gggctccctg
     1381 cggctgaacg tggtatacac ggaagaccac gtgttttctt ctgactatta cagccctctg
     1441 cgggacctgc tgttgaagtc tgcggatgtg gagcccgtgt cagcgtctgc ggcccacatc
     1501 ctgggcgagg tttgccggga gaagcaggag gcggccgtcc cgctggtgcg gctcttccta
     1561 cactatggca gggtggtgcc attcatcagt gccatcgcca gcgcggaggt gaagcggacc
     1621 caggacccca acaccatctt ccgaggaaac tcactggcgt ccaagtgcat tgacgagacc
     1681 atgaagctgg cggggatgca ttacctgcat gtcaccctga agcccgccat cgaggagata
     1741 tgccagagcc acaaaccctg tgaaatcgac cctgtgaagt tgaaagacgg agaaaacctt
     1801 gaaaacaaca tggagaacct acggcagtat gtggaccgcg tcttccacgc catcaccgag
     1861 tctggggtga gctgcccgac cgtcatgtgt gacatcttct tctccctccg ggaggcggcg
     1921 gccaagcgct tccaggatga cccggacgtc aggtacactg cagtgagcag cttcatcttc
     1981 ctgaggttct ttgcgcccgc cattctctcc cccaacctct tccagctcac gccgcaccac
     2041 acggaccccc agacgtccag gacgctgaca ttgatctcca agaccgttca gaccctcggc
     2101 agcctgtcca agtccaaatc tgcgagtttt aaggagtcct acatggctac attttatgaa
     2161 ttcttcaatg agcagaaata tgctgatgcg gtgaagaact tcttggatct gatttcgtcc
     2221 tcggggagaa gagaccccaa gagtgttgag cagcccatcg tgcttaaaga agggttcatg
     2281 atcaagaggg cccaaggacg gaagcgcttt gggatgaaga attttaagaa gagatggttt
     2341 cgcttgacca accatgaatt tacctaccac aaaagcaaag gggaccagcc tctctacagc
     2401 attcccatcg agaacatcct ggcagtggag aagctggagg aggagtcttt caaaatgaaa
     2461 aacatgttcc aggtcatcca gccagagcgt gcgctgtaca tccaggccaa caactgcgtg
     2521 gaggccaagg actggatcga cattctcacc aaagtgagcc agtgcaacca gaagcgcctc
     2581 accgtctacc acccgtccgc ctacctgagc ggccactggc tgtgctgtag ggcgccatcc
     2641 gactcggctc cgggctgctc gccctgcact ggcggcctcc cagccaacat ccagctggac
     2701 attgatgggg accgtgagac ggagcgtatc tactccctct tcaacttgta catgagcaag
     2761 ctggagaaga tgcaggaggc ctgtgggagc aaatctgtgt atgacggccc ggagcaggag
     2821 gagtattcga cgttcgtcat tgacgacccc caggagacct acaagacgct aaagcaagtc
     2881 atcgctgggg ttggggcttt ggagcaggag cacgcccagt ataagaggga caagttcaag
     2941 aagacgaaat atggaagcca ggagcacccc atcggagaca agagcttcca gaactacatc
     3001 cggcagcagt ccgagacctc cactcattcc atttaaggta ccgcgggccc gggatccacc
     3061 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3121 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3181 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3241 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3301 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3361 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3421 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3481 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3541 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3601 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3661 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3721 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3781 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3841 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3901 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3961 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4081 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4141 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4201 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4261 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4321 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4381 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4441 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4501 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4561 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4621 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4681 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4741 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4801 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4861 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4921 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4981 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5041 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5101 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5161 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5221 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5281 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5341 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5401 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5461 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5521 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccatg cat
//
