LOCUS       sequence-432990-        7905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of human AP3B1 (1-677) and AP3M1 AP-3 core 
            hemicomplex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7905)
  AUTHORS   Begley M, Aragon M, Baker RW
  TITLE     A structure-based mechanism for initiation of AP-3 coated vesicle 
            formation.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Dec 24;121(52):e2411974121. doi: 
            10.1073/pnas.2411974121. Epub 2024 Dec 20.
  PUBMED    39705307
REFERENCE   2  (bases 1 to 7905)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-12-24"; volume: "121(52):e2411974121. 
            doi"; pages: " 10.1073/pnas.2411974121. Epub 2024 Dec 20"
COMMENT     Sequence Label: MZB053
FEATURES             Location/Qualifiers
     source          1..7905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             190..212
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2253..2276
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2286..2939
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2897..2919
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     promoter        3072..3090
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3091..3115
                     /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)."
     primer_bind     complement(4474..4492)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4488..4535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4756..5415)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5330..5349
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5416..5518)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5910..5927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6044..6589)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6124..6143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6814..7896)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7858..7877
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(7897..7905,1..69))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
       61 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg cgcgaggcag
      121 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      181 agaaataatt ttgtttaact ttaagaagga gatataccat gagctccaac tcgtttcctt
      241 acaacgaaca gagcggtgga ggggaagcaa cggaattagg tcaagaggcg acgagcacta
      301 tatctccctc tggcgctttt ggcttgttca gttccgatct taagaaaaac gaggacctga
      361 agcaaatgct tgagagcaac aaggattctg ccaaacttga cgccatgaag agaattgttg
      421 ggatgatagc gaaggggaaa aacgcaagtg aactttttcc ggctgttgtc aaaaatgtgg
      481 catccaaaaa catcgaaatt aaaaaacttg tctacgtata cttagtcaga tatgcagagg
      541 agcagcaaga cctggcgctg ctttccatct cgacatttca aagagcactg aaagatccga
      601 accaattaat cagagcatcg gcccttcgtg tgttatctag tatcagagtt cctatcatag
      661 tgcctataat gatgttagcc attaaggaag cctcagctga cctgtccccc tacgtaagaa
      721 agaacgcagc gcatgccatt caaaaactgt actccttgga ccccgaacag aaggaaatgt
      781 tgatagaggt gatagagaag ttacttaagg ataaatcaac tttagtcgcc ggttccgttg
      841 tgatggcttt cgaagaggtc tgcccggacc ggattgactt aattcacaaa aactaccgca
      901 agctttgtaa tttgttagtg gatgtagaag aatggggaca agttgttata attcacatgc
      961 tgacgcgcta tgccagaaca cagttcgtca gtccatggaa ggaaggagat gagcttgagg
     1021 ataacgggaa gaatttttac gagtctgacg acgatcaaaa ggaaaaaaca gataagaaaa
     1081 aaaaacctta tactatggac ccggaccaca gactgttaat acggaacacg aaaccattgt
     1141 tacagtcgag aaacgcggca gttgtgatgg cagttgctca attgtattgg catatttcgc
     1201 ctaaatcaga ggctggaata atctctaaga gcctggtcag acttcttcgc tcaaaccgtg
     1261 aagtacaata tatcgtatta caaaatatcg ccactatgtc catccaacgt aaagggatgt
     1321 tcgaacccta cctgaaatca ttttacgtga gatccacgga cccaactatg ataaaaacct
     1381 taaaacttga gattcttacg aacctggcta atgaagcgaa catttcgacg ttgttacgcg
     1441 aatttcaaac ttacgtcaag tctcaggata aacagtttgc ggctgcgacg attcaaacta
     1501 ttggaagatg tgcaacgaat attcttgagg tcacggacac ttgcctgaac ggtctggtgt
     1561 gccttttgtc taaccgcgac gagatagtcg ttgctgagtc cgttgtagtg atcaagaagc
     1621 ttttacagat gcagcctgcc caacacgggg aaataataaa acacatggcg aagttacttg
     1681 actctatcac cgtgcccgta gctcgcgcaa gtattttatg gttgatcggt gaaaactgtg
     1741 agcgcgtacc caaaatcgca cctgatgtcc tgagaaaaat ggctaaatcg tttacgtcag
     1801 aagatgattt ggttaaatta cagatattaa atttaggggc aaagctttat cttactaatt
     1861 ccaaacaaac taagttgctg actcagtata tcttgaattt agggaaatat gatcaaaatt
     1921 atgacatacg ggatagaacg agattcatta gacagcttat agtccccaac gtgaagtcag
     1981 gtgctttgtc taagtacgct aaaaaaatct tcctggctca gaaaccggcg ccgttactgg
     2041 agagcccatt taaggatcgt gatcactttc aacttggtac actgtctcac actttgaaca
     2101 taaaggcgac tggatacctt gaattgagta attggcccga ggtcgctccg gacccatcgg
     2161 tccggaacgt tgaggtaata gagctggcta aagaatggac tccggcaggc aaagccaagc
     2221 aggagaacag cgctaagaag ttttacagcg gcctggaagt tctgttccag gggcccgggt
     2281 ccggcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
     2341 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
     2401 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
     2461 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
     2521 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
     2581 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
     2641 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
     2701 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
     2761 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
     2821 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
     2881 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
     2941 cggatctggt gccgtaataa tgaggatccg aattcgagct cggcgcgcct gcaggtcgac
     3001 aagcttgcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt acacggccgc
     3061 ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc
     3121 ttagtatatt agttaagtat aagaaggaga tatacatatg attcacagtc ttttcttaat
     3181 caactgttcc ggggatatat ttttggagaa acattggaag agcgtcgtta gtcagtctgt
     3241 ttgtgattac ttttttgagg cccaggagaa agctgccgac gtagaaaacg tacccccagt
     3301 catatcgaca cctcaccact acttgatctc catatataga gataagttat ttttcgtaag
     3361 tgtgatccag actgaagtac caccactgtt cgttatagag ttcttacaca gagtggcgga
     3421 tactttccaa gactatttcg gagaatgctc cgaagccgct ataaaagata acgtagttat
     3481 agtttatgag cttcttgaag agatgttgga taatgggttc ccattggcga ctgaatctaa
     3541 cattctgaag gaattaataa agccccctac aatattgcgc tccgtggtca actctatcac
     3601 cggttcaagt aacgtgggcg atacattgcc aacaggacaa ttaagtaaca tcccttggcg
     3661 gcgcgcgggg gtcaagtata ctaataatga agcttatttc gatgtcgtag aggaaataga
     3721 cgccatcata gacaagagcg gtagcaccgt gtttgccgaa atacaaggag tcattgacgc
     3781 ttgcattaag ctttcgggga tgccagactt gtcattatca tttatgaatc cccgcctttt
     3841 agatgacgtt agctttcatc cttgtatcag atttaagcgt tgggaaagcg agcgcgtctt
     3901 gtctttcatt ccgcccgacg ggaacttccg gttgatttca tatcgggttt cttcgcagaa
     3961 ccttgtagca ataccagtat atgtgaaaca tagtatctcc ttcaaagaga attcgtcttg
     4021 cggccggttc gacattacca ttggacctaa gcaaaacatg gggaaaacaa ttgaaggaat
     4081 aactgtcact gtccacatgc ctaaggttgt acttaatatg aacttaaccc ctacacaagg
     4141 ctcctatacc tttgatcccg tgactaaagt cttaacctgg gatgttggga agataacgcc
     4201 ccaaaaatta ccttctctta agggacttgt gaatttgcaa agcggtgcac ctaaacctga
     4261 agagaaccca agtcttaaca ttcagtttaa gattcagcaa cttgctatca gcggattgaa
     4321 agtaaaccgt ttggacatgt atggtgaaaa atataagcca ttcaaggggg taaagtacgt
     4381 cactaaggca ggtaaattcc aggttcgcac ttagtgagaa ttccgatccg gctgctaaca
     4441 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     4501 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca tttgagaagc
     4561 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     4621 ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc
     4681 attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata
     4741 actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     4801 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     4861 catcagcacc ttgtcgcctt gcgtataata tttgcccata gtgaaaacgg gggcgaagaa
     4921 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     4981 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     5041 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     5101 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     5161 ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag cattcatcag
     5221 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     5281 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     5341 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     5401 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     5461 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     5521 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     5581 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
     5641 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
     5701 ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa
     5761 gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
     5821 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     5881 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     5941 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     6001 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     6061 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     6121 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     6181 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     6241 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     6301 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     6361 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     6421 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     6481 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     6541 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     6601 tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc
     6661 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     6721 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     6781 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6841 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6901 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     6961 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     7021 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     7081 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     7141 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     7201 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     7261 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     7321 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     7381 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     7441 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     7501 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     7561 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     7621 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     7681 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     7741 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     7801 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     7861 atactctgcg acatcgtata acgttactgg tttcacattc accac
//
