LOCUS       sequence-254164-        7893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sec16B as a GFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7893)
  AUTHORS   Budnik A, Heesom KJ, Stephens DJ
  TITLE     Characterization of human Sec16B: indications of specialized, 
            non-redundant functions.
  JOURNAL   Sci Rep. 2011;1:77. doi: 10.1038/srep00077. Epub 2011 Aug 30.
  PUBMED    22355596
REFERENCE   2  (bases 1 to 7893)
  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: "Sci Rep."; 
            date: "2011"; volume: "1:77. doi"; pages: " 10.1038/srep00077. Epub 
            2011 Aug 30"
COMMENT     Sequence Label: pEGFP-Sec16B
FEATURES             Location/Qualifiers
     source          1..7893
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4681..4802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4718..4737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4772..4791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4809..5264)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4946..4967)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5158..5177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5291..5395
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5397..5754
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5397..5588
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5420..5440)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5605..5740
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5789..6583
                     /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             5789..6583
                     /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(5843..5862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6453..6472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6771..6790)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6815..6862
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7191..7779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7680..7699
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctatggaa ctttgggctc cccagaggct gccccagaca
     1381 cgagggaagg ccacagcacc ctcaaaggat ccagaccgag ggtttcggag agatggacat
     1441 catcggcctg tccctcactc ttggcacaat ggagagaggt ttcaccaatg gcaagacaac
     1501 cgtgggagcc cccagccaca gcaggagccc agggcagacc atcagcagca gccccattat
     1561 gcatccaggc caggggactg gcatcagcct gtgtctggag ttgactatta cgaaggtggt
     1621 tatcgcaatc agttgtattc aaggccaggt tatgagaatt catatcagag ctatcagtct
     1681 cccacaatga gggaggaata tgcttatgga agttattact atcatggaca cccacagtgg
     1741 ctgcgggaag aaagagtgcc aaggcaacgg agtccttata tctggcacga agattaccga
     1801 gagcaaaagt accttgatga acatcattat gaaaaccagc acagtccatt tggaacaaat
     1861 agtgagaccc acttccaatc taacagtagg aacccttgta aagacagccc tgcttccaac
     1921 tctggacagg agtggccggg ggagctgttt ccagggagcc tgcttgctga ggcccagaaa
     1981 aataagccat cattggctag tgagtccaac cttctccagc agcgtgagtc tggtctcagc
     2041 tccagcagct atgagctcag tcagtacatc agagatgccc cggagcggga tgatccccca
     2101 gcttcagcag cttggagtcc agttcaggct gaagatgtct cctcagctgg tcccaaagca
     2161 cccatgaagt tctacatccc tcatgttcct gtgagtttcg ggccaggagg tcagctggtg
     2221 catgtaggtc ccagctctcc cactgacggg caagcagccc ttgttgaact gcacagcatg
     2281 gaggttattc ttaatgattc cgaagagcaa gaggagatga gaagtttctc aggacccttg
     2341 attagggaag atgtacataa ggtggatatt atgacgtttt gccagcagaa agcagctcag
     2401 agctgcaaat ctgagacact ggggagcaga gactcagctc tactgtggca gctcttggtt
     2461 ctcctttgtc gccagaatgg gtccatggtg gggtctgaca tcgctgagct gctaatgcaa
     2521 gactgcaaga agctggagaa gtacaagcgg cagccccctg tggccaacct catcaacctg
     2581 accgatgagg actggccagt gctgagctct gggaccccga acctgctcac gggagagatc
     2641 ccccccagtg tggagacacc tgcgcagatc gtggagaaat tcactaggct gctctactat
     2701 ggaaggaaga aggaagcctt ggagtgggcc atgaagaacc acttgtgggg ccatgctttg
     2761 ttcctgtcca gcaagatgga cccacagacc tacagctggg tcatgagtgg cttcaccagc
     2821 acgctggcgc tcaatgaccc actgcagacc ctcttccagc tcatgtcggg gaggattcca
     2881 caggcaacca cgtgttgtgg agaaaagcag tggggagact ggaggcctca cttggctgtg
     2941 attctgtcga atcaggctgg ggacccagag ctgtatcagc gtgcgattgt tgccattggg
     3001 gacaccctgg ctgggaaggg gcttgtggag gcagctcact tctgctatct catggctcac
     3061 gtgccctttg gccactacac cgtgaagaca gaccatctgg tcttgctggg cagcagccac
     3121 agtcaagagt ttttgaaatt tgcaacaact gaggcaatcc agaggacgga aatcttcgag
     3181 tactgtcaga tgctgggccg ccccaaatcc ttcatccctt ctttccaggt gtataagctc
     3241 ctttatgctt cccgtctggc agattatggc ctcgtgtccc aggctttgca ttactgcgaa
     3301 gccattggtg cagctgtctt gagccaggga gagagcagtc accctgtgct attagtggaa
     3361 ctcatcaagc tagcagagaa actgaagctg tcagatcctc tggttttaga aagacgcagt
     3421 ggagacaggg acctggagcc agattggcta gcgcaacttc ggaggcagct ggagcaaaag
     3481 gtagcaggag acattgggga tcctcatcct actcgctcag atatttcggg agccggagga
     3541 acaacaacag aaaacacttt ctaccaggac ttttctggat gtcaaggcta ctctgaagcc
     3601 cctgggtacc gctcagctct gtggctgaca cctgagcaga cctgcctgct ccagcccagc
     3661 ccacagcagc cctttcccct ccagccgggc tcctacccag caggaggggg tgcagggcag
     3721 acagggacac cgaggccttt ttactcagtt cctgagaccc atctaccagg gactggcagc
     3781 agcgtggcag tgacagaggc cactggagga acagtctggg aggaaatgct gcagacacac
     3841 ctgggccctg gagagaacac agtgtctcaa gaaacttccc atcctcctga tggccaagag
     3901 gtcatttcca aaccacagac accattggct gctagagcac gaagtatttc tgagagttcc
     3961 gccaattcag ccaaggagga tgagaaggag tcctctgatg aggctgataa aaactctccc
     4021 cgaaatactg cccagagagg caagctcgga gatgggaagg agcatacaaa gagctcaggg
     4081 tttggctggt tcagctggtt tcgatcgaag cccaccaaga acgcatcccc ctctggagac
     4141 gaggactcct cagacagccc tgactctgag gagaccccca gagcatcttc tccccaccag
     4201 gctggcctgg gcctctcact gacaccttcc cctgagtccc cacctctgcc ggatgttagt
     4261 gccttctcca ggggcagagg tgggggtgaa ggccgaggat ccgcatccag cgggggagca
     4321 gctgcaggtg ctggggttgg aggcttgtct ggaccagaga gtgtttcctt tgagctctgc
     4381 tccaaccctg gtgttcttct tcctccacct gccttaaaag gggctgttcc tctttacaac
     4441 ccatctcagg tgcctcagct gcccacggcc actagcctga atcggccaaa tcgcctagcg
     4501 cagcgtcgct atcccaccca gccatgctga ctcgacggta ccgcgggccc gggatccacc
     4561 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     4621 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4681 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4741 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4801 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4861 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4921 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4981 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5041 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5101 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5161 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5221 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     5281 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5341 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     5401 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     5461 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     5521 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     5581 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5641 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5701 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     5761 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5821 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5881 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5941 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     6001 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6061 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6121 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6181 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6241 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6301 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6361 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6421 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6481 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6541 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6601 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6661 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6721 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     6781 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     6841 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     6901 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     6961 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     7021 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     7081 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7141 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7201 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7261 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7321 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7381 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7441 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7501 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7561 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7621 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7681 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7741 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7801 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7861 gattctgtgg ataaccgtat taccgccatg cat
//
