LOCUS       sequence-335804-        6313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mEmerald-tagged human ERGIC-53 at high level in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6313)
  AUTHORS   Yan R, Chen K, Wang B, Xu K
  TITLE     SURF4-induced tubular ERGIC selectively expedites ER-to-Golgi 
            transport.
  JOURNAL   Dev Cell. 2022 Feb 28;57(4):512-525.e8. doi: 
            10.1016/j.devcel.2021.12.018. Epub 2022 Jan 19.
  PUBMED    35051356
REFERENCE   2  (bases 1 to 6313)
  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"; doi: 
            "10.1016/j.devcel.2021.12.018"; journalName: "Dev Cell"; date: 
            "2022-02-28- 28"; volume: "57"; issue: "4"; pages: "512-525.e8"
COMMENT     Sequence Label: pmEmerald-ERGIC-53
FEATURES             Location/Qualifiers
     source          1..6313
                     /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      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(609..611,612..614,615..1325)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(654..675)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(915..934)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1262..1283
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3041..3162
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3078..3097)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3132..3151
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3169..3624)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3306..3327)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3518..3537
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3651..3755
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3757..4114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3757..3948
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3780..3800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3965..4100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4149..4943
                     /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             4149..4943
                     /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(4203..4222)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4813..4832
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5131..5150)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5175..5222
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5551..6139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6040..6059
                     /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 gctagcgcca
      541 ccatgctgct atccgtgccg ttgctgctcg gcctcctcgg cctggccgtc gccgaaccgg
      601 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      661 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      721 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      781 ggcccaccct cgtgaccacc ttgacctacg gcgtgcagtg cttcgcccgc taccccgacc
      841 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      901 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      961 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1021 tggggcacaa gctggagtac aactacaaca gccacaaggt ctatatcacc gccgacaagc
     1081 agaagaacgg catcaaggtg aacttcaaga cccgccacaa catcgaggac ggcagcgtgc
     1141 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1201 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1261 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1321 acaagtccgg agacgagctg tacaagggtg gcggtggctc gggcggtggt gggtcgggtg
     1381 gcggcggatc tatcgatgaa ttgaattcac gcgttcgcga gttaacggat ccagacggcg
     1441 tgggaggaga ccccgcggtc gcgttgccac atcgccgttt cgagtacaaa tacagcttca
     1501 aggggccgca cctggtgcag agcgacggga ccgtgccctt ctgggcccac gcggggaatg
     1561 ctattccaag ttcagatcaa attcgagtag caccatcttt aaaaagccaa agaggctcag
     1621 tgtggacaaa gacaaaagcg gcctttgaga actgggaagt tgaggtgaca tttcgagtga
     1681 ctggaagagg tcgaattgga gctgatggcc tagcaatttg gtatgcagaa aatcaaggct
     1741 tggagggccc tgtgtttgga tcagctgatc tgtggaatgg tgttggaata ttttttgatt
     1801 cttttgacaa tgatggaaag aaaaataatc ctgctatagt aattataggc aacaatggac
     1861 aaatccatta tgaccatcaa aatgacgggg ctagtcaagc tttggcaagt tgccagaggg
     1921 acttccgcaa caaaccctat cctgtccgag caaagattac ctattaccag aacacactga
     1981 cagtaatgat caataatggc tttacaccag ataaaaatga ttatgaattt tgtgccaaag
     2041 tggaaaatat gattatccct gcacaagggc attttggaat atctgctgca actggaggtc
     2101 ttgcagatga ccatgatgtc ctttcttttc tgactttcca gttgactgaa cctggaaaag
     2161 agccgcccac accagataaa gaaatttcgg aaaaggaaaa agaaaagtat caggaggaat
     2221 ttgagcactt tcaacaagaa ttggataaaa aaaaagagga attccagaag ggccaccccg
     2281 acctccaagg gcagcctgcg gaggaaatat ttgagagtgt aggagatcga gagctaagac
     2341 aagtctttga aggacagaat cgtattcatc ttgaaatcaa gcagctgaac cggcagttag
     2401 atatgattct tgatgaacag agaagatatg tctcttcctt aacagaggaa atctctaaaa
     2461 gaggagcagg aatgcctggg cagcatgggc agattactca acaagaactg gatactgttg
     2521 tgaaaactca gcatgagatt ctgagacaag taaatgaaat gaaaaattcc ttgagtgaaa
     2581 ccgtcagact ggtcagtgga atgcagcacc ctggctctgc tggaggcgtc tatgagacaa
     2641 cacagcactt cattgacatc aaagagcacc tgcacatagt aaagagggac atagataact
     2701 tagtgcagcg aaatatgcca tcaaatgaaa agccgaaatg cccagaacta ccaccatttc
     2761 catcatgttt gtctacggtc cacttcatta tatttgttgt ggtgcaaact gtattattca
     2821 ttggttatat catgtatagg tctcagcaag aagcagctgc caaaaaattc ttttgaggat
     2881 ccctcgagcg gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat
     2941 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3001 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3061 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3121 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3181 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3241 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3301 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3361 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3421 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3481 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3541 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3601 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3661 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3721 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     3781 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3841 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3961 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4021 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4081 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4141 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4201 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4261 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4321 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4381 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4441 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4501 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4561 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4621 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4681 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4741 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4801 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4861 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4921 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4981 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5041 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5101 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5161 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5221 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5281 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5341 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5401 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5461 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5521 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5581 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5641 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5701 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5761 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5821 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5881 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5941 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6001 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6061 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6121 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6181 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6241 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     6301 atatggagtt ccg
//
