LOCUS       sequence-263346-        6654 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 6654)
  AUTHORS   Di Paolo G, Pellegrini L, Letinic K, Cestra G, Zoncu R, Voronov S, 
            Chang S, Guo J, Wenk MR, De Camilli P
  TITLE     Recruitment and regulation of phosphatidylinositol phosphate kinase 
            type 1 gamma by the FERM domain of talin.
  JOURNAL   Nature. 2002 Nov 7. 420(6911):85-9.
  PUBMED    12422219
REFERENCE   2  (bases 1 to 6654)
  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: "Nature. 
            2002 Nov 7. 420(6911):85-9."
COMMENT     Sequence Label: GFP-PIPK1 gamma 87
FEATURES             Location/Qualifiers
     source          1..6654
                     /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    3442..3563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3479..3498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3533..3552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3570..4025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3707..3728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3919..3938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4052..4156
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4158..4515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4158..4349
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4181..4201)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4366..4501
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4550..5344
                     /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             4550..5344
                     /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(4604..4623)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5214..5233
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5532..5551)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5576..5623
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5952..6540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6441..6460
                     /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 ccggccggac tcagatctcg agctcaagct tcgaattcat ggagctggag
     1381 gtaccggacg aggcggagag cgctgaggcg ggggccgtgc cctcggaggc ggcgtgggcg
     1441 gcagagagcg gggcggcggc aggtttggct cagaagaagg cggccccaac agaggttctg
     1501 tccatgacgg cacagccggg ccctggccat gggaagaagt tgggccatcg aggtgtggac
     1561 gcatccggcg aaaccaccta caagaagacc acctcctcca ccctgaaggg tgccatccag
     1621 ctgggcatcg gctacaccgt gggccacctg agctccaagc ccgaacgcga cgtgctcatg
     1681 caggacttct acgtggtgga gagcatcttc ttccccagcg aaggcagcaa cctcaccccc
     1741 gcccaccact tccaggactt ccgcttcaag acctatgcac ctgtcgcctt ccgctacttc
     1801 cgggagctct ttgggatccg gccagatgat tacttgtact ccctgtgcaa tgagccgctg
     1861 atcgagctgt ccaacccggg cgccagtggc tccctcttct acgtcaccag cgacgacgag
     1921 ttcatcatca agaccgtcat gcacaaggag gccgagttcc tgcagaagct gctccctggc
     1981 tactacatga acctcaacca gaacccgcgg acgctgctgc ccaagttcta tgggctgtac
     2041 tgcgtgcagt cggggggcaa gaacatccgc gtcgtggtca tgaacaacat cctgccccgc
     2101 gtggtcaaga tgcacctcaa gttcgacctc aagggctcca cctacaagcg gcgcgccagc
     2161 aagaaggaga aggagaagag cttccccacc tacaaggacc tggacttcat gcaggacatg
     2221 cccgaggggc tcctgctgga cgccgacacc ttcagcgccc tggtcaagac gctgcagcgg
     2281 gactgcctgg tcctggaaag tttcaagatc atggactaca gcctgctgct gggcgtgcac
     2341 aacatcgacc agcacgagcg cgagcggcag gcgcagggcg cccagagcac ctcagatgag
     2401 aagcggcctg tgggccagaa ggcgctctac tccacggcca tggagtccat ccagggtggc
     2461 gccgcgcgcg gggaggccat cgaatcggat gacacgatgg gcgggatccc cgctgtgaac
     2521 ggccgcgggg agcggctgct gctgcacatt ggcatcatcg acatcctgca gtcctacagg
     2581 ttcatcaaga aactggagca cacctggaag gccctcgtcc acgatgggga cacggtgtcc
     2641 gtccaccgcc ccagcttcta tgccgagcgc tttttcaagt tcatgagcaa cacggtcttt
     2701 cggaagaact cctccctgaa gtcctcgccc tccaagaagg ggcgcggcgg agccttgcta
     2761 gctgtgaaac cgctggggcc caccgctgcc ttctcggcca gccagatccc tagcgagcgg
     2821 gaggaggccc agtacgacct gcggggggcc cgcagctacc ccacgctgga ggacgaaggc
     2881 cggcccgacc tcctgccctg cacgccacct tctttcgaag aagccactac agcctccatt
     2941 gccacgactc tgtcatccac atccctctcc attcctgagc ggtccccctc ggagacgtcg
     3001 gagcagccgc ggtacaggcg gcgcacacag tcgtctggac aggatggcag gccgcaggag
     3061 gagccacccg cggaagagga tctgcagcag attacagtgc aggtggagcc tgcgtgcagc
     3121 gtggagattg tggtccccaa agaggaggac gcaggggtgg aggcttcccc ggccggtgcc
     3181 tctgctgctg ttgaagtaga aactgccagc caggcctcag acgaggaggg cgcacctgcc
     3241 agccaggcct cggacgagga ggacgcgccc gccaccgaca tctacttttg agtcgacggt
     3301 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     3361 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3421 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3481 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3541 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     3601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4021 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4081 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4141 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4321 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4441 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4501 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4561 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4621 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4681 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4741 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4801 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4861 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4921 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4981 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5041 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5101 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5161 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5221 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5281 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5341 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5401 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5461 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5521 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5581 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5641 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5701 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5761 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5821 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5881 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5941 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6001 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6061 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6121 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6181 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6241 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6301 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6361 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6421 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6481 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6541 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6601 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
