LOCUS       sequence-364715-        6850 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 6850)
  AUTHORS   Erdmann KS, Mao Y, McCrea HJ, Zoncu R, Lee S, Paradise S, Modregger 
            J, Biemesderfer D, Toomre D, De Camilli P
  TITLE     A role of the Lowe syndrome protein OCRL in early steps of the 
            endocytic pathway.
  JOURNAL   Dev Cell. 2007 Sep . 13(3):377-90.
  PUBMED    17765681
REFERENCE   2  (bases 1 to 6850)
  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: "Dev Cell. 
            2007 Sep . 13(3):377-90."
COMMENT     Sequence Label: pEGFPC1-human APPL1
FEATURES             Location/Qualifiers
     source          1..6850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3578..3699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3615..3634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3669..3688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3706..4161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3843..3864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4055..4074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4188..4292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4294..4651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4294..4485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4317..4337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4502..4637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4686..5480
                     /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             4686..5480
                     /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(4740..4759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5350..5369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5668..5687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5712..5759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6088..6676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6577..6596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagtg ccggggatcg acaagctgcc catcgaggag
     1321 acgctggagg acagcccgca gacaaggtct ttactaggtg tatttgaaga agatgccaca
     1381 gctatttcca actatatgaa ccagttgtat caagctatgc atcggattta tgatgcacag
     1441 aatgaattaa gtgcagcaac acacctgacc tcaaaacttt taaaagaata tgaaaaacag
     1501 cgttttccat tgggaggtga tgatgaagtt atgagctcta cattgcaaca gttttcaaaa
     1561 gttatagatg agcttagctc ttgtcatgca gtgctttcaa ctcaacttgc tgatgccatg
     1621 atgttcccca ttacccagtt taaagaaaga gatctgaaag aaatactaac attaaaggaa
     1681 gtatttcaga ttgcaagtaa tgatcatgat gctgcgatta atagatatag ccgtttatca
     1741 aaaaaaagag aaaatgacaa ggtgaagtat gaagtaacag aagatgtgta cacatccaga
     1801 aagaaacaac accagaccat gatgcattat ttttgtgcat taaatactct tcagtacaag
     1861 aagaaaatag cattgttaga acctctactt gggtacatgc aagctcagat aagtttcttt
     1921 aagatgggtt ctgaaaatct taatgaacaa ctggaagaat ttttagctaa tattggaaca
     1981 agcgttcaga atgttcgcag ggaaatggac agtgatatag agaccatgca acagacaata
     2041 gaggatttgg aagtagccag tgatccctta tatgtgcctg acccagaccc caccaaattt
     2101 cctgttaatc gaaatttaac ccgaaaggct ggatacctta atgctaggaa taaaacaggc
     2161 ttggtgtcat ctacctggga cagacagttt tacttcacgc agggtggaaa tttaatgagt
     2221 caggcccgtg gggatgtagc aggaggcctg gccatggaca tagacaactg ttcagtgatg
     2281 gctgtggact gtgaagacag acgatattgt tttcagatca cctctttcga tggaaaaaaa
     2341 tcttcaattt tgcaagcaga gagtaaaaaa gatcatgaag agtggatctg tacaataaac
     2401 aacatatcta aacaaatata cttaagtgaa aatccagagg aaactgctgc acgagtaaat
     2461 caatcagctc tggaagctgt cactccttcc ccatctttcc agcagaggca cgagagcctg
     2521 cggccagcag caggacaatc tcggccaccg acagctcgaa ccagcagttc aggatcctta
     2581 ggatctgagt ctacaaattt ggctgccctc tctctagatt ctcttgttgc cccagacacc
     2641 ccaatacagt ttgacataat ttctcctgtg tgtgaagatc agcctggcca ggcaaaagcc
     2701 tttggccagg gaggcaggcg tacaaatcca tttggagaat ctggaggaag tacaaaatct
     2761 gaaactgaag attctattct tcatcagtta tttattgtcc gattccttgg ttcaatggag
     2821 gtgaaatcag atgaccatcc agatgttgtt tatgaaacta tgcgccaaat cttagctgcc
     2881 cgggccatcc ataacatctt tcgtatgaca gaatcgcatt tattagtcac ttgtgactgt
     2941 ttaaagttaa ttgatccaca gacacaagtt acaaggctca cgtttccatt accttgtgta
     3001 gttttgtatg ctacacacca ggaaaataag cgcctttttg gatttgttct tcggacatca
     3061 agcgggagaa gtgaaagtaa tctgtcatca gtctgctata tatttgagtc aaacaatgag
     3121 ggggaaaaga tatgtgattc tgttggactg gcaaaacaga tagctttgca tgctgaactg
     3181 gatcgtaggg catcagaaaa acaaaaagaa atagagagag taaaagagaa gcaacagaaa
     3241 gaactcaata aacaaaaaca gattgaaaag gacttggaag aacaaagtcg gttgatagct
     3301 gcttccagta gaccaaacca agccagtagt gaggggcagt ttgttgtcct tagcagtagc
     3361 cagtcagaag agagtgattt gggagaagga ggaaagaaga gagaatcaga agcataagaa
     3421 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     3481 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3541 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3721 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3781 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3841 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3901 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3961 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4021 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4081 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4141 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4261 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4621 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4681 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4741 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4801 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4861 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4921 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4981 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5041 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5101 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5161 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5221 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5281 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5341 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5401 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5461 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5521 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5581 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5641 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5701 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5761 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5821 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5881 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5941 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6001 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6721 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6781 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6841 tggagttccg
//
