LOCUS       sequence-73784-e        5367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP-Rab4AS22N (dominant negative mutant) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5367)
  AUTHORS   Rzomp KA, Moorhead AR, Scidmore MA
  TITLE     The GTPase Rab4 interacts with Chlamydia trachomatis inclusion 
            membrane protein CT229.
  JOURNAL   Infect Immun. 2006 Sep;74(9):5362-73.
  PUBMED    16926431
REFERENCE   2  (bases 1 to 5367)
  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: "Infect 
            Immun."; date: "2006-09"; volume: "74(9)"; pages: "5362-73"
COMMENT     Sequence Label: EGFP-Rab4AS22N
FEATURES             Location/Qualifiers
     source          1..5367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    816..937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(853..872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     907..926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(944..1399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1081..1102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1293..1312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1426..1530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1532..1889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1532..1723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1555..1575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1740..1875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1924..2718
                     /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             1924..2718
                     /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(1978..1997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2588..2607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2906..2925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2950..2997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3326..3914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3815..3834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4089..4392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4393..4596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4547..4567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4635..4644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4641..4643,4644..4646,4647..5357)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4641..4643,4644..4646,4647..5357)
                     /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(4686..4707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4947..4966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5294..5315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatccaccat gtcgcagacg gccatgtccg aaacctacga ttttttgttt aagttcttgg
       61 ttattggaaa tgcaggaact ggcaaaaatt gcttacttca tcagtttatt gaaaaaaaat
      121 tcaaagatga ctcaaatcat acaataggag tggaatttgg ttcaaagata ataaatgttg
      181 gtggtaaata tgtaaagtta caaatatggg atacagcagg acaagaacga ttcaggtccg
      241 tgacgagaag ttattaccga ggcgcggccg gggctctcct cgtctatgat atcaccagcc
      301 gagaaaccta caatgcgctt actaattggt taacagatgc ccgaatgcta gcgagccaga
      361 acattgtgat catcctttgt ggaaacaaga aggacctgga tgcagatcgt gaagttacct
      421 tcttagaagc ctccagattt gctcaagaaa atgagctgat gtttttggaa acaagtgcgc
      481 tcacagggga gaatgtagaa gaggcttttg tacagtgtgc aagaaaaata cttaacaaaa
      541 tcgaatcagg tgagctggac ccagaaagaa tgggctcagg tattcagtac ggagatgctg
      601 ccttgagaca gctgaggtca ccgcggcgcg cacaggcccc gaacgctcag gagtgtggtt
      661 gttagctcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
      721 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
      781 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
      841 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      901 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
      961 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1021 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1081 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1141 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1201 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1261 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1321 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1381 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1501 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     1561 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1621 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1681 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     1741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1801 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     1861 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     1921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     1981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2101 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     2461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2701 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     2761 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     2821 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     2881 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     2941 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3001 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3061 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3121 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     3181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3721 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3961 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4021 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4081 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4141 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4201 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4261 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4321 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4381 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4441 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4501 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4561 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     4621 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     4681 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4741 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     4801 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     4861 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     4921 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4981 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     5041 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     5101 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     5161 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     5221 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     5281 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5341 tggacgagct gtacaagtcc ggactca
//
