LOCUS       sequence-331747-        8553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Sec31 tagged with HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8553)
  AUTHORS   Weigel AV, Chang CL, Shtengel G, Xu CS, Hoffman DP, Freeman M, Iyer 
            N, Aaron J, Khuon S, Bogovic J, Qiu W, Hess HF, Lippincott-Schwartz 
            J
  TITLE     ER-to-Golgi protein delivery through an interwoven, tubular network 
            extending from ER.
  JOURNAL   Cell. 2021 Apr 29;184(9):2412-2429.e16. doi: 
            10.1016/j.cell.2021.03.035. Epub 2021 Apr 13.
  PUBMED    33852913
REFERENCE   2  (bases 1 to 8553)
  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.cell.2021.03.035"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2412-2429.e16"
COMMENT     Sequence Label: Halo-Sec31
FEATURES             Location/Qualifiers
     source          1..8553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        159..462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        463..666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     617..637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      705..714
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             711..1601
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    5439..5560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5476..5495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5530..5549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5567..6022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5704..5725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5916..5935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6049..6153
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6155..6512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6155..6346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6178..6198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6363..6498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6547..7341
                     /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             6547..7341
                     /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(6601..6620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7211..7230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7529..7548)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7573..7620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7949..8537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8438..8457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
       61 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
      121 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      181 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      241 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      301 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      361 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      421 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      481 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      541 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      601 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      661 agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggtcgccacc atggcagaaa
      721 tcggtactgg ctttccattc gacccccatt atgtggaagt cctgggcgag cgcatgcact
      781 acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt cctgcacggt aacccgacct
      841 cctcctacgt gtggcgcaac atcatcccgc atgttgcacc gacccatcgc tgcattgctc
      901 cagacctgat cggtatgggc aaatccgaca aaccagacct gggttatttc ttcgacgacc
      961 acgtccgctt catggatgcc ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca
     1021 ttcacgactg gggctccgct ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca
     1081 aaggtattgc atttatggag ttcatccgcc ctatcccgac ctgggacgaa tggccagaat
     1141 ttgcccgcga gaccttccag gccttccgca ccaccgacgt cggccgcaag ctgatcatcg
     1201 atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag
     1261 tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc
     1321 gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag
     1381 aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg ggcaccccag
     1441 gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct aactgcaagg
     1501 ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac ctgatcggca
     1561 gcgagatcgc gcgctggctg tcgacgctcg agatttccgg catgtacaag tccggactca
     1621 gatctcgagc tcaagcttcg aattcgatga agttaaagga agtagatcgt acagccatgc
     1681 aggcatggag ccctgcccag aatcacccca tttacctagc aacaggaaca tctgctcagc
     1741 aattggatgc aacatttagt acgaatgctt cccttgagat atttgaatta gacctctctg
     1801 atccatcctt ggatatgaaa tcttgtgcca cattctcctc ttctcacagg taccacaagt
     1861 tgatttgggg gccttataaa atggattcca aaggagatgt ctctggagtt ctgattgcag
     1921 gtggtgaaaa tggaaatatt attctctatg atccttctaa aattatagct ggagacaagg
     1981 aagttgtgat tgcccagaat gacaagcata ctggcccagt gagagccttg gatgtgaaca
     2041 ttttccagac taatctggta gcttctggtg ctaatgaatc tgaaatctac atatgggatc
     2101 taaataattt tgcaacccca atgacaccag gagccaaaac acagccgcca gaagatatca
     2161 gctgcattgc atggaacaga caagttcagc atattttagc atcagccagt cccagtggcc
     2221 gggccactgt atgggatctt aggaaaaatg agccaatcat caaagtcagt gaccatagta
     2281 acagaatgca ttgttctggg ttggcatggc atcctgatgt tgctactcag atggtccttg
     2341 cctccgagga tgaccggtta ccagtgatcc agatgtggga tcttcgattt gcttcctctc
     2401 cacttcgtgt cctggaaaac catgccaggg ggattttggc aattgcttgg agcatggcag
     2461 atcctgaatt gttactgagc tgtggaaaag atgctaagat tctctgctcc aatccaaaca
     2521 caggagaggt gttatatgaa cttcccacca acacacagtg gtgcttcgat attcagtggt
     2581 gtccccgaaa tcctgctgtc ttatcagctg cttcgtttga tgggcgtatc agtgtttatt
     2641 ctatcatggg aggaagcaca gatggtttaa gacagaaaca agttgacaag ctttcatcat
     2701 cttttgggaa tcttgatccc tttggcacag gacagcccct tcctccgtta caaattccac
     2761 agcagactgc tcagcatagt atagtgctgc ctctgaagaa gccgcccaag tggattcgaa
     2821 ggcctgttgg tgcttctttt tcatttggag gcaaactggt tacgtttgag aatgtcagaa
     2881 tgccttctca tcagggagct gagcagcagc agcagcagca ccatgtgttc attagtcagg
     2941 ttgtaacaga aaaggagttc ctcagccgat cagaccaact tcagcaggct gtgcagtcac
     3001 aaggatttat caattattgc caaaaaaaaa ttgatgcttc tcagactgaa tttgagaaaa
     3061 atgtgtggtc ctttttgaag gtaaactttg aggatgattc tcgtggaaaa taccttgaac
     3121 ttctaggata cagaaaagaa gatctaggaa agaagattgc tttggccttg aacaaagtgg
     3181 atggagccaa tgtggctctt aaagactctg accaagtagc acagagtgat ggggaggaga
     3241 gccctgctgc tgaagagcag ctcttgggag agcacattaa agaggaaaaa gaagaatctg
     3301 aatttctacc ctcatctgga ggaacattta atatctctgt cagtggggac attgatggtt
     3361 taattactca ggctttgctg acgggcaatt ttgagagtgc tgttgacctt tgtttacatg
     3421 ataaccgcat ggccgatgcc attatattgg ccatagcagg tggacaagaa ctcttggctc
     3481 gaacccagaa aaaatacttc gcaaaatccc aaagcaaaat taccaggctc atcactgcag
     3541 tggtgatgaa gaactggaaa gagattgttg agtcttgtga tcttaaaaat tggagagagg
     3601 ctttagctgc agtattgact tatgcaaagc cggatgaatt ttcagccctt tgtgatcttt
     3661 tgggaaccag gcttgaaaat gaaggagata gcctcctgca gactcaagca tgtctctgct
     3721 atatttgtgc ggggaatgta gagaaattag ttgcatgttg gactaaagct caagatggaa
     3781 gccacccttt gtcacttcag gatctgattg agaaagttgt catcctgcga aaagctgtgc
     3841 aactcactca agccatggac actagtactg taggagttct cttggctgcg aagatgagtc
     3901 agtatgccaa tttgttggca gctcagggca gtattgctgc agccttggct tttcttcctg
     3961 acaacaccaa ccagccaaat atcatgcagc ttcgtgacag actttgtaga gcacaaggag
     4021 agcctgtagc aggacatgaa tcacctaaaa ttccgtacga gaaacagcag ctccccaagg
     4081 gcaggcctgg accagttgct ggccaccacc agatgccaag agttcaaact caacaatatt
     4141 atccccatgg agaaaatcct ccacctccgg gtttcataat gcatggaaat gttaatccaa
     4201 atgctgctgg tcagcttccc acatctccag gtcatatgca cacccaggta ccaccttatc
     4261 cacagccaca gccttatcaa ccagcccagc cgtatccctt cggaacaggg gggtcagcaa
     4321 tgtatcgacc tcagcagcct gttgctcctc ctacttcaaa cgcttaccct aacacccctt
     4381 acatatcttc tgcttcttcc tatactgggc agtctcagct gtacgcagca cagcaccagg
     4441 cctcttcacc tacctccagc cctgctactt ctttccctcc tcccccttcc tctggagcat
     4501 ccttccagca tggcggacca ggagctccac catcatcttc agcttatgca ctgcctcctg
     4561 gaacaacagg tacactgcct gctgccagtg agctgcctgc gtcccaaaga acaggtcctc
     4621 agaatggttg gaatgaccct ccagctttga acagagtacc caaaaagaag aagatgcctg
     4681 aaaacttcat gcctcctgtt cccatcacat caccaatcat gaacccgttg ggtgaccccc
     4741 agtcacaaat gctgcagcaa cagccttcag ctccagtacc actgtcaagc cagtcttcat
     4801 tcccacagcc acatcttcca ggtggccagc ccttccatgg cgtacagcaa cctcttggtc
     4861 aaacaggcat gccaccatct ttttcaaagc ccaatattga aggtgcccca ggggctccta
     4921 ttggaaatac cttccagcat gtgcagtctt tgccaacaaa aaaaattacc aagaaaccta
     4981 ttccagatga gcacctcatt ctaaagacca catttgagga tcttattcag cgctgccttt
     5041 cttcagcaac agaccctcaa accaagagga agctagatga tgccagcaaa cgtttggagt
     5101 ttctgtatga taaacttagg gaacagacac tttcaccaac aatcaccagt ggtttacaca
     5161 acattgcaag gagcattgaa actcgaaact actcagaagg attgaccatg catacccaca
     5221 tagttagcac cagcaacttc agtgagacct ctgctttcat gccagttctc aaagttgttc
     5281 tcacccaggc caataagctg ggtgtctaag gatccaccgg atctagataa ctgatcataa
     5341 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     5401 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     5461 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5521 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     5581 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5641 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5701 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5761 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5821 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5881 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5941 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6001 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6061 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6121 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     6181 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6241 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6301 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6361 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6421 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6481 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     6541 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6601 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6661 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6721 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6781 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6841 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6901 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6961 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7021 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     7081 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7141 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7201 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7261 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7321 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     7381 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     7441 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     7501 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     7561 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     7621 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     7681 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     7741 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     7801 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7861 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7921 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7981 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     8041 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     8101 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     8161 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     8221 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     8281 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     8341 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     8401 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     8461 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     8521 ggcggagcct atggaaaaac gccagcaacg cgg
//
