LOCUS       sequence-273547-        6148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Eukaryotic expression of rat GRASP55 full-length.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  TITLE     Yanzhuang Wang Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6148)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pEGFP-N1-GRASP55
FEATURES             Location/Qualifiers
     source          1..6148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2088..2097
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2094..2096,2097..2099,2100..2813)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2094..2096,2097..2099,2100..2810)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2139..2160)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2400..2419)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2747..2768
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2936..3057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2973..2992)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3027..3046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3064..3519)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3201..3222)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3413..3432
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3546..3650
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3652..4009
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3652..3843
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3675..3695)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3860..3995
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4044..4838
                     /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             4044..4838
                     /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(4098..4117)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4708..4727
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5026..5045)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5070..5117
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5446..6034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5935..5954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcg gccgccactg tgctggatat ctgcagaatt ccaccacact
      661 ggactagtgg atccatgggc tcctcgcaga gcgtcgagat cccgggcggc ggcacagagg
      721 gctaccacgt cctgcgggta caggaaaatt ccccaggaca cagagctgga ctggagcctt
      781 tctttgactt tattgtttct attaatggtt cgagattaaa taaggacaac gacactctga
      841 aggacttgct gaaagccaac gtggagaagc ctgtgaagat gctcatctac agcagcaaaa
      901 ccctggagct gcgggaggcc tcggtcacac ccagcaacct gtggggcggc cagggcttgt
      961 tgggggttag catccggttc tgcagtttcg atggggccaa cgaaaatgtt tggcacgtgc
     1021 tggaagtgga atcgaattcc cccgcagcac tggcggggct gagacctcac agcgattaca
     1081 tcattggagc tgataccgtc atgaatgagt ctgaagacct gttcagcctt attgagaccc
     1141 atgaagcaaa gcctcttaag ctctacgtgt acaacacaga cactgacaac tgccgagaag
     1201 tgatcattac accaaattct gcatggggtg gggaaggcag cctcggctgt ggcatcggct
     1261 atggttattt gcaccgaata cctacgcgtc cctttgaaga aggaaagaag atttcccttc
     1321 caggacaaat gactggcacc cccatgactc ctcctaagga tgggtttacg gaggtccagc
     1381 tgtcctcagt cagcccccca tctttgtcac caccaggaac tgcaggagtt gagcagagtc
     1441 tgtctggact gtccattagc tcagctccac cggctgtcag caatgttctc agtacaggtg
     1501 taccgactgt accattgttg ccgccacaag taaaccagtc ccttgcttct gtgccaccaa
     1561 tgaacccagc tgccacatta ccaagtctga tgcctttgtc agcagggctg cctagcctcc
     1621 ccaacctccc cagcctctcc aacttcaacc tccctgctcc gcacatcatg ccaggggtcg
     1681 gcttgccaga gctcgggaag ccgggtttgc cacctcttcc gtccttgcct ccccgaaact
     1741 tacctggcat tgcacctctc cccatgccgt ccgacttcct cccgtcattc cctttggttc
     1801 cagagggctc ttctgcagcc agcgcagggg aaccactgtc ttcccttcct gccatgggcc
     1861 caccttctga ccctgtcatg actactgcaa aggcagacac ctcctccctc actgtggatg
     1921 tgatgtctcc tgcttccaag gtccccacca ctgttgagga cagagtgagc gactgcaccc
     1981 cagccatgga gaagcctgtg tctgcagtta cggatgcaaa tgcttctgag gcttctaagc
     2041 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga
     2101 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2161 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2221 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2281 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2341 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2401 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2461 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2521 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     2581 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     2641 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2701 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2761 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     2821 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2881 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2941 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3001 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3061 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3121 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3181 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3241 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3301 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3361 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3421 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3481 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3541 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3601 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3661 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3721 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3841 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3901 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3961 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4021 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4081 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4141 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4201 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4261 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4321 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4381 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4441 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4501 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4561 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4621 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4681 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4741 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4801 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4861 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4921 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4981 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5041 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5101 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5161 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5221 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5281 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5341 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5401 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5461 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5521 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5581 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5641 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5701 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5761 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5821 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5881 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5941 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6001 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6061 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6121 tgtggataac cgtattaccg ccatgcat
//
