LOCUS       sequence-210516-        6673 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mAzami-labeled galectin 3 (Gal3) as a marker for endosomal
            leakage or rupture..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6673)
  AUTHORS   Dingjan I, Verboogen DR, Paardekooper LM, Revelo NH, Sittig SP, 
            Visser LJ, Mollard GF, Henriet SS, Figdor CG, Ter Beest M, van den 
            Bogaart G
  TITLE     Lipid peroxidation causes endosomal antigen release for 
            cross-presentation.
  JOURNAL   Sci Rep. 2016 Feb 24;6:22064. doi: 10.1038/srep22064.
  PUBMED    26907999
REFERENCE   2  (bases 1 to 6673)
  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: "2016-02-24"; volume: "6:22064. doi"; pages: " 
            10.1038/srep22064"
COMMENT     Sequence Label: mAzami-Green-Galectin
FEATURES             Location/Qualifiers
     source          1..6673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      972..981
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             978..1655
                     /codon_start=1
                     /product="humanized monomeric Azami-Green fluorescent 
                     protein"
                     /label=hmAzami-Green
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             978..1655
                     /codon_start=1
                     /product="monomeric Azami-Green fluorescent protein 
                     (Karasawa et al., 2003)"
                     /label=mAzami-Green
                     /translation="MVSVIKPEMKIKLCMRGTVNGHNFVIEGEGKGNPYEGTQILDLNV
                     TEGAPLPFAYDILTTVFQYGNRAFTKYPADIQDYFKQTFPEGYHWERSMTYEDQGICTA
                     TSNISMRGDCFFYDIRFDGTNFPPNGPVMQKKTLKWEPSTEKMYVEDGVLKGDVNMRLL
                     LEGGGHYRCDFKTTYKAKKEVRLPDAHKIDHRIEILKHDKDYNKVKLYENAVARYSMLP
                     SQAK"
     CDS             2544..2573
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2589..2606
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2629..2646)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2635..2859
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2905..3333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2992..3011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3202..3223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3342..3362)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3347..3676
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3527..3662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3589..3608
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3724..3771
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3790..4188
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4346..4467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4383..4402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4437..4456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4516..4532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4529..4551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4540..4556
                     /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(4564..4570,4571..4588,4589..4594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4609..4630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4747..4764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4918..5506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4998..5017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5677..6468,6469..6537))
                     /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     6300..6319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6538..6642)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcca
      961 gcacagtggc ggccgccatg gtgagcgtga tcaagcccga gatgaagatc aagctgtgca
     1021 tgaggggcac cgtgaacggc cacaacttcg tgatcgaggg cgagggcaag ggcaacccct
     1081 acgagggcac ccagatcctg gacctgaacg tgaccgaggg cgcccccctg cccttcgcct
     1141 acgacatcct gaccaccgtg ttccagtacg gcaacagggc cttcaccaag taccccgccg
     1201 acatccagga ctacttcaag cagaccttcc ccgagggcta ccactgggag aggagcatga
     1261 cctacgagga ccagggcatc tgcaccgcca ccagcaacat cagcatgagg ggcgactgct
     1321 tcttctacga catcaggttc gacggcacca acttcccccc caacggcccc gtgatgcaga
     1381 agaagaccct gaagtgggag cccagcaccg agaagatgta cgtggaggac ggcgtgctga
     1441 agggcgacgt gaacatgagg ctgctgctgg agggcggcgg ccactacagg tgcgacttca
     1501 agaccaccta caaggccaag aaggaggtga ggctgcccga cgcccacaag atcgaccaca
     1561 ggatcgagat cctgaagcac gacaaggact acaacaaggt gaagctgtac gagaacgccg
     1621 tggccaggta ctccatgctg cccagccagg ccaagggagg aggaggatct gcagacaatt
     1681 tttcgctcca tgatgcgtta tctgggtctg gaaacccaaa ccctcaagga tggcctggcg
     1741 catgggggaa ccagcctgct ggggcagggg gctacccagg ggcttcctat cctggggcct
     1801 accccgggca ggcaccccca ggggcttatc ctggacaggc acctccaggc gcctaccctg
     1861 gagcacctgg agcttatccc ggagcacctg cacctggagt ctacccaggg ccacccagcg
     1921 gccctggggc ctacccatct tctggacagc caagtgccac cggagcctac cctgccactg
     1981 gcccctatgg cgcccctgct gggccactga ttgtgcctta taacctgcct ttgcctgggg
     2041 gagtggtgcc tcgcatgctg ataacaattc tgggcacggt gaagcccaat gcaaacagaa
     2101 ttgctttaga tttccaaaga gggaatgatg ttgccttcca ctttaaccca cgcttcaatg
     2161 agaacaacag gagagtcatt gtttgcaata caaagctgga taataactgg ggaagggaag
     2221 aaagacagtc ggttttccca tttgaaagtg ggaaaccatt caaaatacaa gtactggttg
     2281 aacctgacca cttcaaggtt gcagtgaatg atgctcactt gttgcagtac aatcatcggg
     2341 ttaaaaaact caatgaaatc agcaaactgg gaatttctgg tgacatagac ctcaccagtg
     2401 cttcatatac catgatataa ggatccctcc ccccccccta acgttactgg ccgaagccgc
     2461 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     2521 ggcaatgtga gggcccgcgg ttcgaacaaa aactcatctc agaagaggat ctgaatatgc
     2581 ataccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     2641 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2701 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2761 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2821 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2881 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3121 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3241 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3301 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3361 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3421 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3481 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3541 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3601 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3661 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     3721 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     3781 aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     3841 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     3901 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     3961 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     4021 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     4081 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     4141 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc acttcgtggc
     4201 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     4261 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4321 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4381 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4441 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4501 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4561 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4621 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4681 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6661 gccacctgac gtc
//
