LOCUS       sequence-334997-        5323 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 5323)
  AUTHORS   Allaire PD, Seyed Sadr M, Chaineau M, Seyed Sadr E, Konefal S, 
            Fotouhi M, Maret D, Ritter B, Del Maestro RF, McPherson PS
  TITLE     Interplay between Rab35 and Arf6 controls cargo recycling to 
            coordinate cell adhesion and migration.
  JOURNAL   J Cell Sci. 2013 Feb 1;126(Pt 3):722-31. doi: 10.1242/jcs.112375. 
            Epub 2012 Dec 21.
  PUBMED    23264734
REFERENCE   2  (bases 1 to 5323)
  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 
            Sci."; date: "2013-02-1"; volume: "126(Pt 3):722-31. doi"; pages: " 
            10.1242/jcs.112375. Epub 2012 Dec 21"
COMMENT     Sequence Label: GFP-Rab35 WT
FEATURES             Location/Qualifiers
     source          1..5323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      644..653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(650..652,653..655,656..1366)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(650..652,653..655,656..1366)
                     /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(695..716)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(956..975)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1303..1324
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2148..2269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2185..2204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2239..2258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2276..2731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2413..2434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2625..2644
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2758..2862
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2864..3221
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2864..3055
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2887..2907)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3072..3207
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3256..4050
                     /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             3256..4050
                     /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(3310..3329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3920..3939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4238..4257)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4282..4329
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4658..5246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5147..5166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa
      661 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      721 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      781 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      841 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      901 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      961 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1021 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1081 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1141 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1201 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1261 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1321 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     1381 catggcccgg gactacgacc acctcttcaa gctgctcatc atcggcgaca gcggtgtggg
     1441 caagagcagt ttactgttgc gttttgcaga caacactttc tcaggcagct acatcaccac
     1501 gatcggagtg gatttcaaga tccggaccgt ggagatcaac ggggagaagg tgaagctgca
     1561 gatctgggac acagcggggc aggagcgctt ccgcaccatc acctccacgt attatcgggg
     1621 gacccacggg gtcattgtgg tttacgacgt caccagtgcc gagtcctttg tcaacgtcaa
     1681 gcggtggctt cacgaaatca accagaactg tgatgatgtg tgccgaatat tagtgggtaa
     1741 taagaatgac gaccctgagc ggaaggtggt ggagacggaa gatgcctaca aattcgccgg
     1801 gcagatgggc atccagttgt tcgagaccag cgccaaggag aatgtcaacg tggaagagat
     1861 gttcaactgc atcacggagc tggtcctccg agcaaagaaa gacaacctgg caaaacagca
     1921 gcagcaacaa cagaacgatg tggtgaagct cacgaagaac agtaaacgaa agaaacgctg
     1981 ctgctaagaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
     2041 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2101 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2701 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2881 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2941 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3061 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3121 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3181 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3421 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3781 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4081 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4201 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4261 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4321 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4381 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4441 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4501 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5221 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5281 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atc
//
