LOCUS       sequence-331274-        6173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed mannII-BFP simultaneously with a RUSH Hook Str-KDEL..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6173)
  AUTHORS   McCaughey J, Stevenson NL, Mantell JM, Neal CR, Paterson A, Heesom 
            K, Stephens DJ
  TITLE     A general role for MIA3/TANGO1 in secretory pathway organization and
            function.
  JOURNAL   J Cell Sci. 2021 Aug 5. pii: 271201. doi: 10.1242/jcs.259075.
  PUBMED    34350936
REFERENCE   2  (bases 1 to 6173)
  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.
            2021 Aug 5. pii: 271201. doi: 10.1242/jcs.259075."
COMMENT     Sequence Label: StrKDEL-IRES-mannosidase II-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..6173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(51..625,626..628,629..637)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(218..235)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     445..464
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1026..1724
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1897..2018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2120..2136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2120..2136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2133..2155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2144..2160
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2168..2174,2175..2192,2193..2198))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2213..2234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2351..2368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2522..3110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2602..2621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3281..4072,4073..4141))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3904..3923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4142..4246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4321..4340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4512..4891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4892..5095
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5046..5066
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5140..5159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5140..5158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          5923..6152
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     6135..6154
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc cccctctccc
       61 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
      121 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
      181 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
      241 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
      301 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
      361 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
      421 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
      481 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
      541 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
      601 ttcctttgaa aaacacgatg ataagcttgc cacaacccgg gaggcgcgcc atgaagttaa
      661 gtcgccagtt caccgtgttt ggcagcgcga tcttctgcgt cgtaatcttc tcactctacc
      721 tgatgctgga caggggtcac ttggactacc ctcggggccc gcgccaggag ggctcctttc
      781 cgcagggcca gctttcaata ttgcaagaaa agattgacca tttggagcgt ttgctcgctg
      841 agaacaacga gattatctca aatatcagag actcagtcat caacctgagc gagtctgtgg
      901 aggacggccc gcgggggtca ccaggcaacg ccagccaagg ctccatccac ctccactcgc
      961 cacagttggc cctgcaggct gaccccagag actgtttgga tcccaccggt cgccaccgga
     1021 attccatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1081 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1141 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1201 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     1261 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     1321 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     1381 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     1441 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
     1501 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacatcaag accacatata
     1561 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     1621 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     1681 ccagatactg cgacctccct agcaaactgg ggcacaagct taatggggcc ctcgaggcct
     1741 taaggccggc cttaaggcct cgagggccaa ttaattaact ctagataact gatcataatc
     1801 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1861 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1921 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1981 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg cattctagtt
     2041 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     2101 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2161 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2221 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2281 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2341 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2401 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2461 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2521 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2581 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2641 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2701 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2761 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2821 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2881 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2941 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3001 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3061 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4261 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     4321 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     4381 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     4441 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     4501 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4561 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4621 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4681 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4741 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4801 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4861 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4921 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4981 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5041 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     5101 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     5161 gacccaagct tggtaccgag ctcggatcga tatctgcggc ctagctagcg cttaaggcct
     5221 gttaaccggt atggatgtat gcgtccgtct tgccctgtgg ctcctctggg gactcctcct
     5281 gcaccagggc cagagcctca gccatagtca cagtgagaag gcgacaggaa ccagctcggg
     5341 cgccggttta aacgacccta gcaaagactc aaaagctcag gtgtccgctg ccgaggctgg
     5401 cattactgga acatggtaca atcagctcgg gagcaccttt attgtgactg ctggagccga
     5461 tggagccctc accggaacat acgaatctgc tgtgggaaac gccgaatcac ggtacgtcct
     5521 cactggccga tacgatagtg cccctgccac cgacggatct gggactgccc tgggatggac
     5581 tgtcgcttgg aaaaacaact accggaatgc tcattctgcc acaacatgga gtggacagta
     5641 cgtgggaggc gctgaggcta gaatcaatac acagtggctg ctcacatctg gcacaaccga
     5701 ggcaaatgct tggaaatcca ccctggtggg acatgacaca ttcaccaaag tgaaaccctc
     5761 cgccgcttca atcgatgccg ccaaaaaagc cggagtcaac aacggcaatc ctctggatgc
     5821 cgtccagcag ctcggaccga aagatgaact gtgagcggcc gcatagataa ctgatccagt
     5881 gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac tccctctcaa
     5941 aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat
     6001 tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca gaaaagacaa
     6061 tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac acttgagtga
     6121 caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aac
//
