LOCUS       sequence-73742-e        5376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP-Rab4B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5376)
  AUTHORS   Rzomp KA, Scholtes LD, Briggs BJ, Whittaker GR, Scidmore MA
  TITLE     Rab GTPases are recruited to chlamydial inclusions in both a 
            species-dependent and species-independent manner.
  JOURNAL   Infect Immun. 2003 Oct;71(10):5855-70.
  PUBMED    14500507
REFERENCE   2  (bases 1 to 5376)
  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: "2003-10"; volume: "71(10)"; pages: "5855-70"
COMMENT     Sequence Label: EGFP-Rab4B
FEATURES             Location/Qualifiers
     source          1..5376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    801..922
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(838..857)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     892..911
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(929..1384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1066..1087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1278..1297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1411..1515
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1517..1874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1517..1708
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1540..1560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1725..1860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1909..2703
                     /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             1909..2703
                     /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(1963..1982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2573..2592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2891..2910)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2935..2982
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3311..3899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3800..3819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4074..4377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4378..4581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4532..4552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4620..4629
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4626..4628,4629..4631,4632..5342)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4626..4628,4629..4631,4632..5342)
                     /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(4671..4692)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4932..4951)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5279..5300
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aattcaccat ggctgagacc tacgacttcc tcttcaaatt cctggtgatt ggcagtgcag
       61 gaactggcaa atcatgtctc cttcatcagt tcattgagaa taagttcaaa caggactcca
      121 accacacaat cggcgtggag tttggatccc gggtggtcaa cgtgggtggg aagactgtga
      181 agctacagat ttgggacacg gctggccagg agcggtttcg gtcagtgacg cggagttatt
      241 accgaggggc ggctggagcc ctgctggtgt acgacatcac cagccgggag acatacaact
      301 cactggctgc ctggctgacg gatgcccgca ccctggccag ccccaacatc gtggtcatcc
      361 tctgtggcaa caagaaggac ctggaccctg agcgggaggt cactttcctg gaggcctccc
      421 gctttgccca ggagaatgag ctgatgttcc tggagaccag cgctctcaca ggcgagaacg
      481 tggaggaggc gttcctcaag tgtgcccgca ctatcctcaa caagattgac tcaggcgagc
      541 tagacccgga gaggatgggc tctggcattc agtacgggga tgcgtccctc cgccagcttc
      601 ggcagcctcg gagtgcccag gccgtggccc ctcagccgtg tggctgctga ctcgacggta
      661 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
      721 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
      781 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
      841 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      901 ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc
      961 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1021 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1081 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1141 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1201 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1261 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1321 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1381 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1441 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1501 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     1561 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1621 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1681 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1741 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1801 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1861 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     1921 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     1981 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2041 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     2101 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2161 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2221 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2281 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2341 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2401 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     2461 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2521 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2581 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2641 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2701 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2761 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2821 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     2881 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     2941 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3001 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3061 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3121 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     3181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3421 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     4021 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4081 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     4141 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     4201 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     4261 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     4321 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     4381 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     4441 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     4501 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     4561 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
     4621 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4681 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     4741 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     4801 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     4861 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     4921 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     4981 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5041 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5101 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5161 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5221 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5281 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5341 agtccggccg gactcagatc tcgagctcaa gcttcg
//
