LOCUS       sequence-366309-        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Endoplasmic Reticulum marker GFP-ER [TM(SAC1)] in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Alpy F, Rousseau A, Schwab Y, Legueux F, Stoll I, Wendling C, 
            Spiegelhalter C, Kessler P, Mathelin C, Rio MC, Levine TP, Tomasetto
            C
  TITLE     STARD3 or STARD3NL and VAP form a novel molecular tether between 
            late endosomes and the ER.
  JOURNAL   J Cell Sci. 2013 Dec 1;126(Pt 23):5500-12. doi: 10.1242/jcs.139295. 
            Epub 2013 Oct 8.
  PUBMED    24105263
REFERENCE   2  (bases 1 to 7959)
  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: "J Cell 
            Sci."; date: "2013-12-1"; volume: "126(Pt 23):5500-12. doi"; pages: 
            " 10.1242/jcs.139295. Epub 2013 Oct 8"
COMMENT     Sequence Label: pQCXIP EGFP-ER [TM(SAC1)]
FEATURES             Location/Qualifiers
     source          1..7959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             156..572
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     502..524
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     540..556
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        609..912
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        913..1116
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1113..1137
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1262..1271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1268..1270,1271..1273,1274..1984)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1268..1270,1271..1273,1274..1984)
                     /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(1313..1334)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1574..1593)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1921..1942
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(2276..2850,2851..2853,2854..2862)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2443..2460)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2670..2689
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2894..3292
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3670..4095
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4198..4220)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4356..4376)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4361..4690
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4541..4676
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4712..4731
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4812..4829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4983..5571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5063..5082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5742..6533,6534..6602))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6365..6384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6603..6707)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6775..6793
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        6893..7272
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7273..7476
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             7477..7652
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    7715..7914
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt
       61 ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag catcgttctg tgttgtctct
      121 gtctgactgt gtttctgtat ttgtctgaaa attagggcca gactgttacc actcccttaa
      181 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
      241 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
      301 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
      361 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
      421 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
      481 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
      541 tttatccagc cctcactcct tctctaggcg ccggaattga agatctgggg gatcgatcct
      601 ctagagtccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      661 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      721 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      781 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      841 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      901 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      961 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1021 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1081 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     1141 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
     1201 cgcggccggg aacggtgcat tggaacggac ctgcaggaat tgatccgcgg ccgcaccggt
     1261 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1321 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1381 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1441 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1501 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1561 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1621 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1681 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1741 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1801 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1861 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1921 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1981 caagtccgga ctcagatctc gagctcaagc ttcgaattcc cgggtcttcc tggctttgcc
     2041 tgttatcatg gttgttgcct tttcaatgtg cattatctgt ttgcttatgg ctggtgacac
     2101 ttggacagaa acactggcct atgtgctctt ctggggagtt gcaagcattg gaacattttt
     2161 tatcattctt tacaatggca aagattttgt cgatgctccc agactggtcc agaaagaaaa
     2221 gatagactga ggtaccgcgg gcccgggatc cggaattccg cccccccccc ccccccccct
     2281 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2341 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2401 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2461 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2521 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2581 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2641 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2701 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     2761 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2821 tggttttcct ttgaaaaaca cgatgataag cttgccacaa cccacaagga gacgaccaat
     2881 tcggaactaa accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
     2941 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
     3001 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
     3061 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
     3121 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
     3181 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
     3241 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattgcc
     3301 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tccgaccgaa
     3361 gccacccggg gcggccccgc cgaccccgca cccgcccccg aggcccaccg actctagtcg
     3421 agggctgcag cgctgcagag gccgagtgca gaactgctcc aaagggacct caaggctttc
     3481 cgagggacac taggctgact ccatcgagcc agtgtagaga taagcttatc gattagtcca
     3541 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca
     3601 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa
     3661 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     3721 ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac
     3781 agatggaaca gggtcgaccc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     3841 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     3901 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     3961 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     4021 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     4081 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
     4141 acccaccacc gggaggtaag ctggctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4201 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4261 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     4321 gtcacgtagc gatagcggag tgtagatccg gctgtggaat gtgtgtcagt tagggtgtgg
     4381 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4441 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4501 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4561 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4621 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4681 cttttgcaaa aagcttactg gcttaactat gcggcatcag agcagattgt actgagagtg
     4741 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     4801 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4861 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4921 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4981 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5041 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5101 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5161 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5221 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5281 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5341 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5401 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5461 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5521 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5581 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5641 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5701 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5761 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5821 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5881 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5941 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6001 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     6061 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6121 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6181 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6241 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6301 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     6361 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6421 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6481 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6541 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6601 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6661 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6721 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6781 cgtatcacga ggccctttcg tcttcaagaa ttagcttggc cattgcatac gttgtatcca
     6841 tatcataata tgtacattta tattggctca tgtccaacat taccgccatg ttgacattga
     6901 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     6961 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7021 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7081 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7141 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7201 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     7261 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     7321 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     7381 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     7441 aggcgtgtac ggtgggaggt ctatataagc agagctcaat aaaagagccc acaacccctc
     7501 actcggcgcg ccagtcttcc gatagactgc gtcgcccggg tacccgtatt cccaataaag
     7561 cctcttgctg tttgcatccg aatcgtggtc tcgctgttcc ttgggagggt ctcctctgag
     7621 tgattgacta cccacgacgg gggtctttca tttgggggct cgtccgggat ttggagaccc
     7681 ctgcccaggg accaccgacc caccaccggg aggtaagctg gccagcaact tatctgtgtc
     7741 tgtccgattg tctagtgtct atgtttgatg ttatgcgcct gcgtctgtac tagttagcta
     7801 actagctctg tatctggcgg acccgtggtg gaactgacga gttctgaaca cccggccgca
     7861 accctgggag acgtcccagg gactttgggg gccgtttttg tggcccgacc tgaggaaggg
     7921 agtcgatgtg gaatccgacc ccgtcaggat atgtggttc
//
