LOCUS       sequence-487633-        6733 bp    DNA     circular SYN 01-OCT-2025
DEFINITION  Rab effector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 6733)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: GFP Slyp4a
FEATURES             Location/Qualifiers
     source          1..6733
                     /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    3461..3582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3498..3517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3552..3571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3589..4044)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3726..3747)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3938..3957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4071..4175
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4177..4534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4177..4368
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4200..4220)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4385..4520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4569..5363
                     /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             4569..5363
                     /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(4623..4642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5233..5252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5551..5570)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5595..5642
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5971..6559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6460..6479
                     /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 atctatgtcg gagttactgg acctttcttt tctgtctgag
     1321 gaggaaaagg atttgattct cagtgttcta cagcgagatg aagaggtccg gaaagcagat
     1381 gagaaaagga ttaggcgact aaagaatgag ttactggaga taaaaaggaa aggggccaag
     1441 aggggcagcc aacactacag tgatcggacc tgtgcccggt gccaggagag cctgggccgt
     1501 ttgagtccca aaaccaatac ttgtcggggt tgtaatcacc tggtgtgtcg ggactgccgc
     1561 atacaggaaa gcaatggtac ctggaggtgc aaggtgtgcg ccaaggaaat agagttgaag
     1621 aaagcaactg gggactggtt ttatgaccag aaagtgaatc gctttgctta ccgcacaggt
     1681 agtgagataa tcaggatgtc cctgcgccac aaacctgcag tgagtaaaag agagacagtg
     1741 ggacagtccc tccttcatca gacacagatg ggtgacatct ggccaggaag aaagatcatt
     1801 caggagcggc agaaggagcc cagtgtgcta tttgaagtgc caaagctgaa aagtggaaag
     1861 agtgcattgg aagctgagag tgagagtctg gatagcttca cagctgactc ggatagcacc
     1921 tccaggagag actctctgga taaatctggc ctctttccag aatggaagaa gatgtctgct
     1981 cccaaatctc aagtagaaaa ggaaactcag cctggaggtc aaaatgtggt atttgtggat
     2041 gagggtgaga tgatatttaa gaagaacacc agaaaaatcc tcaggccttc agagtacact
     2101 aaatctgtga tagatcttcg cccagaagat gtggtacatg aaagtggctc cttgggagac
     2161 agaagcaaat ccgtcccagg cctcaatgtg gatatggaag aggaagaaga agaagaagac
     2221 attgaccacc tagtgaagtt acatcgccag aagctagcca gaagcagcat gcaaagtggc
     2281 tcctccatga gtacgatcgg cagcatgatg agcatctaca gtgaagctgg tgatttcggg
     2341 aacatctttg tgactggcag gattgccttt tccctgaagt atgagcagca aacccagagt
     2401 ctggttgtcc atgtgaagga gtgccatcag ctggcctatg ctgatgaagc caagaagcgc
     2461 tctaacccat atgtgaagac ttaccttctg cctgacaagt cccgccaagg aaaaagaaaa
     2521 accagcatca agcgggacac tgttaatcca ctatatgatg agacgctgag gtatgagatc
     2581 ccagaatctc tcctggccca gaggaccctg cagttctcag tttggcatca tggtcgtttt
     2641 ggcagaaaca ctttccttgg agaggcagag atccagatgg attcctggaa gcttgataag
     2701 aaactggatc attgcctccc tttacatgga aagatcagtg ctgagtcccc gactggcttg
     2761 ccatcacaca aaggcgagtt ggtggtttca ttgaaataca tcccagcctc caaaacccct
     2821 gttggaggtg accggaaaaa gagtaaaggt ggggaagggg gagagctcca ggtgtggatc
     2881 aaagaagcca agaacttgac ggctgccaaa gcaggaggga cttcagacag ctttgtcaag
     2941 ggatacctcc ttcccatgag gaacaaggcc agtaaacgta aaactcctgt gatgaagaag
     3001 accctgaatc ctcactacaa ccatacattt gtctacaatg gtgtgaggct ggaagatcta
     3061 cagcatatgt gcctggaact gactgtgtgg gaccgggagc ccctggccag caatgacttc
     3121 ctgggagggg tcaggctggg tgttggcact gggatcagta atggggaagt ggtggactgg
     3181 atggactcga ctggggaaga agtgagcctg tggcagaaga tgcgacagta cccagggtct
     3241 tgggcagaag ggactctgca gctccgttcc tcaatggcca agcagaagct gggtttatga
     3301 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     3361 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3421 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3481 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3541 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3601 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3661 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3721 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3781 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3841 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3901 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3961 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4021 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4201 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4261 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4321 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4381 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4441 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4501 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4561 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4621 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4681 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4741 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4801 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4861 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4921 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4981 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5041 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5101 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5161 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5221 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5281 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5341 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5401 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5461 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5521 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5581 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5641 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5701 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5761 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5821 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5881 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5941 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6001 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6061 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6121 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6181 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6241 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6301 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6361 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6421 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6481 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6601 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6661 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6721 atatggagtt ccg
//
