LOCUS       sequence-9837-ex        5229 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 5229)
  AUTHORS   Botelho RJ, Teruel M, Dierckman R, Anderson R, Wells A, York JD, 
            Meyer T, Grinstein S
  TITLE     Localized biphasic changes in phosphatidylinositol-4,5-bisphosphate 
            at sites of phagocytosis.
  JOURNAL   J Cell Biol. 2000 Dec 25. 151(7):1353-68.
  PUBMED    11134066
REFERENCE   2  (bases 1 to 5229)
  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: "J Cell 
            Biol. 2000 Dec 25. 151(7):1353-68."
COMMENT     Sequence Label: CFP-C1-PLCdelta-PH
FEATURES             Location/Qualifiers
     source          1..5229
                     /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"
     CDS             join(598..600,601..603,604..1314)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(643..664)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(904..923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1251..1272
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1315..1371
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2017..2138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2054..2073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2108..2127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2145..2600)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2282..2303)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2494..2513
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2627..2731
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2733..3090
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2733..2924
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2756..2776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2941..3076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3125..3919
                     /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             3125..3919
                     /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(3179..3198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3789..3808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4107..4126)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4151..4198
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4527..5115
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5016..5035
                     /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 gctagcgatg
      601 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      661 gacgtaaacg gccacaggtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      721 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      781 gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      841 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgtac catcttcttc
      901 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      961 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1021 ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1081 atcaaggccc acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1141 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1201 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1261 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagagatct
     1321 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc catggactcg
     1381 ggccgggact tcctgaccct gcacggccta caggatgatg aggatctaca ggcgctgctg
     1441 aagggcagcc agctcctgaa ggtgaagtcc agctcatgga ggagagagcg cttctacaag
     1501 ttgcaggagg actgcaagac catctggcag gagtcccgca aggtcatgcg gaccccggag
     1561 tcccagctgt tctccatcga ggacattcag gaggtgcgaa tggggcaccg cacggagggt
     1621 ctggagaagt tcgcccgtga tgtgcccgag gaccgctgct tctccattgt cttcaaggac
     1681 cagcgcaata cactagacct catcgcccca tcgccagctg atgcccagca ctgggtgctg
     1741 gggctgcaca agatcatcca ccactcaggc tccatggacc agcgtcagaa gctacagcac
     1801 tggattcact cctgcttgcg aaaagctgac aaaaacaagg acaacaagat gagcttcaag
     1861 gagctgcaga acttcctgaa ggagctcaac atccagtaat ctagataact gatcataatc
     1921 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1981 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2041 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2101 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2161 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2221 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2281 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2341 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2401 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2461 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2521 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2581 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2701 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2761 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2821 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2881 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2941 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3001 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3061 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3121 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3181 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3241 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3301 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3361 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3421 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3481 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3541 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3601 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3661 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3721 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3781 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3841 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3901 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4081 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4141 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4201 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4261 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4321 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4381 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4441 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4501 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4561 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4621 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     4681 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4741 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4801 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4861 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4921 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4981 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5041 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5101 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5161 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5221 gccatgcat
//
