LOCUS       sequence-8329-ex        7916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7916)
  AUTHORS   Bhattacharyya D, Glick BS
  TITLE     Two mammalian Sec16 homologues have nonredundant functions in 
            endoplasmic reticulum (ER) export and transitional ER organization.
  JOURNAL   Mol Biol Cell. 2007 Mar;18(3):839-49. doi: 10.1091/mbc.e06-08-0707
  PUBMED    17192411
REFERENCE   2  (bases 1 to 7916)
  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: "10.1091/mbc"; 
            journalName: "Mol Biol Cell"; date: "2007-03"; volume: "18"; issue: 
            "3"; pages: "839-49"
COMMENT     Sequence Label: pmGFP-Sec16S
FEATURES             Location/Qualifiers
     source          1..7916
                     /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 with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     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    4704..4825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4741..4760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4795..4814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4832..5287)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4969..4990)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5181..5200
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5314..5418
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5420..5777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5420..5611
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5443..5463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5628..5763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5812..6606
                     /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             5812..6606
                     /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(5866..5885)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6476..6495
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6794..6813)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6838..6885
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7214..7802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7703..7722
                     /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 aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagga atggaacttt gggctcccca gaggctgccc
     1381 cagacacgag ggaaggccac agcaccctca aaggatccag accgagggtt tcggagagat
     1441 ggacatcatc ggcctgtccc tcactcttgg cacaatggag agaggtttca ccaatggcaa
     1501 gacaaccgtg ggagccccca gccacagcag gagcccaggg cagaccatca gcagcagccc
     1561 cattatgcat ccaggccagg ggactggcat cagcctgtgt ctggagttga ctattacgaa
     1621 ggtggttatc gcaatcagtt gtattcaagg ccaggttatg agaattcata tcagagctat
     1681 cagtctccca caatgaggga ggaatatgct tatggaagtt attactatca tggacaccct
     1741 cagtggctgc aggaagaaag agtgccaagg caacggagtc cttatatctg gcacgaagat
     1801 taccgagagc aaaagtacct tgatgaacat cattatgaaa accagcacag tccatttgga
     1861 acaaatagtg agacccactt ccaatctaac agtaggaacc cttgtaaaga cagccctgct
     1921 tccaactctg gacgggagtg gccgggggag ctgtttccag ggagcctgct tgctgaggcc
     1981 cagaaaaata agccatcatt ggctagtgag tccaaccttc tccagcagcg tgagtctggt
     2041 ctcagctcca gcagctatga gctcagtcag tacatcagag atgccccgga gcgggatgat
     2101 cccccagctt cagcagcttg gagtccagtt caggctgatg tctcctcagc tggtcccaaa
     2161 gcacccatga agttctacat ccctcatgtt cctgtgagtt tcgggccagg aggtcagctg
     2221 gtgcgtgtag gtcccagctc tcccactgac gggcaagcag cccttgttga attgcacagc
     2281 atggaggtta ttcttaatga ttccgaagag caagaggaga tgagaagttt ctcaggaccc
     2341 ttgattaggg aagatgtaca taaggtggat attatgacgt tttgccagca gaaagcagct
     2401 cagagctgca aatctgagac actggggagc agagactcag ctctactgtg gcagctcttg
     2461 gttctccttt gtcgccagaa tgggtccatg gtggggtctg acatcgctga gctgctaatg
     2521 caagactgca agaagctgga gaagtacaag cggcagcccc ctgtggccaa cctcatcaac
     2581 ctgaccgatg aggactggcc agtgctgagc tctgggaccc cgaacctgct cacgggagag
     2641 atccccccca gtgtggagac acctgcgcag atcgtggaga aattcactag gctgctctac
     2701 tatggaagga agaaggaagc cttggagtgg gccatgaaga accacttgtg gggccatgct
     2761 ttgttcctgt ccagcaagat ggacccacag acctacagct gggtcatgag tggcttcacc
     2821 agcacgctgg cgctcaatga cccactgcag accctcttcc agctcatgtc ggggaggatt
     2881 ccacaggcag ccacgtgttg tggagaaaag cagtggggag actggaggcc tcacttggct
     2941 gtgattctgt cgaatcaggc tggggaccca gagctgtatc agcgtgcgat tgttgccatt
     3001 ggggacaccc tggctgggaa ggggcttgtg gaggcagctc acttctgcta tctcatggct
     3061 cacgtgccct ttggccacta caccgtgaag acagaccatc tggtcttgct gggcagcagc
     3121 cacagtcaag agtttttgaa atttgcaaca actgaggcaa tccagaggac ggaaatcttc
     3181 gagtactgtc agatgctggg ccgccccaaa tccttcatcc cttctttcca ggtgtataag
     3241 ctcctttatg cttcccgtct ggcagattat ggcctcgtgt cccaggcttt gcattactgc
     3301 gaagccattg gtgcagctgt cttgagccag ggagagagca gtcaccctgt gctattagtg
     3361 gaactcatca agctagcaga gaaactgaag ctgtcagatc ctctggtttt agaaagacgc
     3421 agtggagaca gggacctgga gccagattgg ctagcgcaac ttcggaggca gctggagcaa
     3481 aaggtagcag gagacattgg ggatcctcat cctactcgct cagatatttc gggagccgga
     3541 ggaacaacaa cagaaaacac tttctaccag gacttttctg gatgtcaagg ctactctgaa
     3601 gcccctgggt accgctcagc tctgtggctg acacctgagc agacctgcct gctccagccc
     3661 agcccacagc agccctttcc cctccagccg ggctcctacc cagcaggagg gggtgcaggg
     3721 cagacaggga caccgaggcc tttttactca gttcctgaga cccatctacc agggactggc
     3781 agcagcgtgg cagtgacaga ggccactgga ggaacagtct gggaggaaat gctgcagaca
     3841 cacctgggcc ctggagagaa cacagtgtct caagaaactt cccagcctcc tgatggccaa
     3901 gaggtcattt ccaaaccaca gacaccattg gctgctagac cacgaagtat ttctgagagt
     3961 tccgccagtt cagccaagga ggatgagaag gagtcctctg atgaggctga taaaaactct
     4021 ccccgaaata ctgcccagag aggcaagctc ggagatggga aggagcatac aaagagctca
     4081 gggtttggct ggttcagctg gtttcgatcg aagcccacca agaacgcatc ccctgctgga
     4141 gacgaggact cctcagacag ccctgactct gaggagaccc ccagagcatc ttctccccac
     4201 caggctggcc tgggcctctc actgacacct tcccctgagt ccccacctct gccggatgtt
     4261 agtgccttct ccaggggcag aggtgggggt gaaggccgag gatccgcatc cagcggggga
     4321 gcagctgcgg gcgctggggt tggaggcttg tccggaccag agagtgtttc ctttgagctc
     4381 tgctccaacc ctggtgttct tcttcctcca cctgccttaa aaggggctgt tcctctttac
     4441 aacccatctc aggtgcctca gctgcccacg gccactagcc tgaatcggcc aaatcgccta
     4501 gctcagcgtc gctatcccac ccagccatgc tgactcaagc ttcgaattct gcagtcgacg
     4561 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     4621 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     4681 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4741 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4801 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     4861 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4921 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4981 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5041 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5101 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5161 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5221 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5281 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5341 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5401 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     5461 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5521 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5581 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5641 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5701 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5761 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5821 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5881 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5941 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     6001 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     6061 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     6121 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     6181 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     6241 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     6301 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     6361 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     6421 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     6481 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6541 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6601 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6781 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6841 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6901 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6961 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     7021 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     7081 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7141 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7201 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7261 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7321 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     7381 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7441 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7501 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7561 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7621 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7681 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7741 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7801 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7861 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
