LOCUS       sequence-73793-e        5337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP-Rab6AT27N (Dominant Negative mutant) in mammalian 
            cells).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5337)
  AUTHORS   Moorhead AR, Rzomp KA, Scidmore MA
  TITLE     The Rab6 effector Bicaudal D1 associates with Chlamydia trachomatis 
            inclusions in a biovar-specific manner.
  JOURNAL   Infect Immun. 2007 Feb;75(2):781-91. Epub 2006 Nov 13.
  PUBMED    17101644
REFERENCE   2  (bases 1 to 5337)
  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: "2007-02"; volume: "75(2)"; pages: "781-91. Epub 2006
            Nov 13"
COMMENT     Sequence Label: EGFP-Rab6AT27N
FEATURES             Location/Qualifiers
     source          1..5337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    786..907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(823..842)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     877..896
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(914..1369)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1051..1072)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1263..1282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1396..1500
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1502..1859
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1502..1693
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1525..1545)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1710..1845
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1894..2688
                     /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             1894..2688
                     /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(1948..1967)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2558..2577
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2876..2895)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2920..2967
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3296..3884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3785..3804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4059..4362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4363..4566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4517..4537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4605..4614
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4611..4613,4614..4616,4617..5327)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4611..4613,4614..4616,4617..5327)
                     /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(4656..4677)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4917..4936)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5264..5285
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gatccaccat gtccacgggc ggagacttcg ggaatccgct gaggaaattc aagctggtgt
       61 tcctggggga gcaaagcgtt ggaaagaact ctttgatcac cagattcatg tatgacagtt
      121 ttgacaacac ctatcaggca acaattggca ttgacttttt atcaaaaact atgtacttgg
      181 aggatcgaac agtacgattg caattatggg acacagcagg tcaagagcgg ttcaggagct
      241 tgattcctag ctacattcgt gactccactg tggcagttgt tgtttatgat atcacaaatg
      301 ttaactcatt ccagcaaact acaaagtgga ttgatgatgt cagaacagaa agaggaagtg
      361 atgttatcat catgctagta ggaaataaaa cagatcttgc tgacaagagg caagtgtcaa
      421 ttgaggaggg agagaggaaa gccaaagagc tgaatgttat gtttattgaa actagtgcaa
      481 aagctggata caatgtaaag cagctctttc gacgtgtagc agcagctttg ccgggaatgg
      541 aaagcacaca ggacagaagc agagaagata tgattgacat aaaactggaa aagcctcagg
      601 agcaaccagt cagtgaagga ggctgttcct gctaactcga cggtaccgcg ggcccgggat
      661 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
      721 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
      781 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      841 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      901 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
      961 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     1021 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     1081 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     1141 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     1201 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     1261 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     1321 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     1381 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1441 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1501 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     1561 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     1621 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     1681 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     1741 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     1801 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     1861 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     1921 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     1981 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2041 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     2101 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2161 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2221 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2281 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2341 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2401 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     2461 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     2521 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2581 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2641 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     2701 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     2761 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     2821 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     2881 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     2941 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     3001 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     3061 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     3121 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     3181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3421 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3961 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     4021 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4081 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4141 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4201 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4261 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4321 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     4381 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4441 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4501 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     4561 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggtgagca
     4621 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4681 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4741 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4801 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     4861 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4921 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4981 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     5041 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     5101 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     5161 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     5221 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     5281 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc ggactca
//
