LOCUS       sequence-365328-        8991 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of clathrin adaptor AP-1 core complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8991)
  AUTHORS   Ren X, Farias GG, Canagarajah BJ, Bonifacino JS, Hurley JH
  TITLE     Structural basis for recruitment and activation of the AP-1 clathrin
            adaptor complex by Arf1.
  JOURNAL   Cell. 2013 Feb 14;152(4):755-67. doi: 10.1016/j.cell.2012.12.042.
  PUBMED    23415225
REFERENCE   2  (bases 1 to 8991)
  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.2012.12"; journalName: "Cell"; date: "2013-02-14- 
            14"; volume: "152"; issue: "4"; pages: "755-67"
COMMENT     Sequence Label: pST44-AP-1 core
FEATURES             Location/Qualifiers
     source          1..8991
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1798..1820
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3611..3631
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3644..4297
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4255..4277
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     RBS             4344..4366
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      4526..4535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5708..5726)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5722..5769
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5836..5927
                     /gene="bla"
                     /label=AmpR promoter
     terminator      6792..6831
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      6862..7450
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7351..7370
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7604..7621
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7636..7775
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7701..7720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7861..7883
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7877..8068)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     8305..8324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8305..8323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             8355..8377
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     8856..8873
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8857..8873
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             8886..8908
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             8928..8945
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(8973..8991,1..2)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 gatccatgac tgactcaaaa tatttcacca cgaccaagaa aggggagatc ttcgagctga
       61 aggcagaact caacagtgac aagaaggaga agaagaagga ggcagtgaag aaagtgattg
      121 catcgatgac cgtgggcaaa gatgtcagtg ccctcttccc cgatgtggtc aactgcatgc
      181 agacggacaa cctggagctg aagaagctgg tatacctcta cttgatgaat tacgccaaga
      241 gtcagcctga catggccatt atggccgtca acacctttgt gaaggactgt gaggacccca
      301 accccctcat ccgagccctg gcagtgcgga ccatgggctg catccgcgtt gacaagatca
      361 cagagtacct gtgcgagcca ctccggaagt gcctgaagga cgaggaccca tatgtgcgca
      421 agacagcagc tgtgtgcgtg gccaagctcc acgacatcaa cgcccagctg gtggaggacc
      481 agggcttcct ggacaccctt aaagacctca tctccgactc taaccccatg gtggtggcca
      541 atgcagtggc agcgctctca gaaattgccg agtctcaccc cagcagcaac ctgctcgatc
      601 tgaacccaca gtccatcaac aagctgctga cagccctcaa tgagtgcacc gagtggggcc
      661 agatcttcat cctggactgc ctcgccaact atatgcccaa ggacgaccgc gaggcccaga
      721 gcatctgtga gcgggtcacc cccaggctct cccatgccaa ctccgctgtg gtgctctctg
      781 ctgtgaaggt gctgatgaag ttcatggaga tgttgtctaa ggacttggac tactacggca
      841 cactgctcaa gaagctggcc ccacccctgg tcacactgct gtcagccgag ccagagctgc
      901 agtatgtggc cctgcgcaac atcaatctca tcgtgcagaa aaggcctgag atcctgaagc
      961 atgagatgaa ggtgttcttc gtgaagtaca acgaccctat ctacgtgaag ctggagaagc
     1021 tggacatcat gatccgcctg gcctctcagg ccaacatcgc ccaggtgttg gcagagctga
     1081 gagagtacgc aacagaagtg gatgtggact ttgtacggaa ggctgtgcgt gctattggcc
     1141 gctgcgccat caaggtggag caatctgcgg agcgctgtgt gagcacgctg ctcgacctca
     1201 tccagaccaa ggtcaactat gtggtccagg aggccatcgt ggtcatcaag gacatcttcc
     1261 gcaagtaccc caacaagtat gagagtgtga ttgccacact gtgtgagaat ctggactccc
     1321 tggatgagcc tgaggcccgg gctgccatga tctggattgt gggcgagtac gcggaacgga
     1381 tcgacaacgc agatgagctg ctggagagct tcctcgaggg cttccatgac aagagcacac
     1441 aggtccagct gcagctgctg acagccattg tgaaactctt tctaaagaag ccaacagaga
     1501 cccaggagct ggtgcagcag gtcctcagtt tggccactca ggactcagat aacccagacc
     1561 tgcgggaccg tggctacatc tactggcgcc tgctgtccac ggacccggtg gcagccaagg
     1621 aggtggtgtt ggctgagaag ccactcatct ctgaagagac ggacctcatt gagcccacac
     1681 tgttagacga gcttatctgc tacatcggca cgctggcttc cgtctaccat aagcctccca
     1741 gtgcctttgt ggaggggtga aagcttcgcc agggttttcc cagtcgagct caataatttt
     1801 gtttaacttt aagaaggaga tatacatgcc agcccccatc agattgcggg agctgatccg
     1861 gaccatccgg acagcccgaa cccaagctga ggaacgagaa atgatccaga aagaatgtgc
     1921 tgcaatccgg tcctcattta gagaagaaga caatacatac cggtgtcgga atgtggcaaa
     1981 gttactatat atgcacatgc tgggctaccc tgctcacttt ggacagttgg agtgcctcaa
     2041 gctaatcgcc tcacaaaaat tcacagacaa acgcattggc tatttaggag caatgctatt
     2101 attagatgaa agacaagatg tgcatcttct catgaccaac tgtatcaaga atgatcttaa
     2161 tcatagcacg cagtttgtgc aggggctagc actttgtacc ctcggctgca tgggctcctc
     2221 ggagatgtgc agagatcttg cgggagaggt agaaaagctt ctgaaaacct ccaactctta
     2281 cttaagaaaa aaggcagcac tgtgtgctgt tcacgtcatc aggaaggttc ctgaacttat
     2341 ggagatgttt ttaccagcaa caaaaaattt attgaatgag aagaaccatg gtgtccttca
     2401 cacgtctgta gtcctcctca cagagatgtg tgaacggagc ccagacatgc tcgcacattt
     2461 cagaaagctt gtgccccaat tagttcgtat tctaaagaac ctcatcatgt ccggatattc
     2521 cccagaacat gatgtctctg gtatcagtga cccctttttg caggtacgaa ttttgcggtt
     2581 attaagaatt ttaggacgga atgacgatga ttcaagtgaa gctatgaatg atatattagc
     2641 acaggttgcc actaatacag agactagtaa aaatgtagga aatgctattc tgtatgagac
     2701 ggttttgact atcatggata ttaaatcaga gagtggattg cgagttctag ccataaatat
     2761 tcttggtcga ttcttattga acaatgacaa aaatattaga tatgttgctc tgacatctct
     2821 gttgaagact gtgcagacag atcataacgc tgtacagagg catagaagca caattgtgga
     2881 ctgtctgaaa gatctggatg tctccataaa aagacgtgca atggaactga gttttgccct
     2941 ggtgaatggg aataatatcc gaggcatgat gaaagaatta ctttattttc tggattcctg
     3001 tgagccagaa tttaaagctg attgtgcatc tggaatcttc cttgctgcag aaaagtacgc
     3061 accttccaaa cggtggcata tagatacaat tatgcgtgtc ttgacaacgg caggaagtta
     3121 tgttcgcgat gatgcagttc ctaacttgat tcagttaata accaacagtg tggagatgca
     3181 cgcttacacc gtccagcgcc tgtacaaggc gattctcggt gactattctc aacaaccctt
     3241 ggtacaagtg gctgcgtggt gtataggtga atatggcgat cttcttgtgt ctggccagtg
     3301 tgaagaggaa gagcctattc aggtgacaga agatgaagtg ctggatattt tagaaagtgt
     3361 cctgatctct aatatgtcca cctctgtgac aagaggctat gctctcactg ccattatgaa
     3421 gctgtctacc cgattcacct gtactgtaaa ccgaattaag aaagtggttt ccatctatgg
     3481 gagcagcatc gacgtagaac tccagcagag ggcagtagag tacaacgcac tttttaagaa
     3541 gtatgaccac atgaggtctg ccctacttga aagaatgcct gtcatggaaa aagtgaccac
     3601 aaatggccct gaaaacctgt attttcaggg cgccatggga tccatgtccc ctatactagg
     3661 ttattggaaa attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga
     3721 aaaatatgaa gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt
     3781 tgaattgggt ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac
     3841 acagtctatg gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc
     3901 aaaagagcgt gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt
     3961 ttcgagaatt gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct
     4021 acctgaaatg ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga
     4081 tcatgtaacc catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga
     4141 cccaatgtgc ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat
     4201 cccacaaatt gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg
     4261 gcaagccacg tttggtggtg gcgaccatcc tccaaaaatc gattaaggta ccagcggata
     4321 acaatttcac atccggaaat aattttgttt aactttaaga aggagatata catatgtccg
     4381 ccagcgccgt ctacgtactg gatctgaagg gcaaggtgct catctgcagg aactaccgtg
     4441 gggatgtgga catgtcagag gtggagcact tcatgcccat tctgatggag aaggaggagg
     4501 agggcatgct gtcacctatc ttggcccatg gtggcgttcg tttcatgtgg attaagcaca
     4561 acaacctgta cctggtcgcc acttcaaaaa agaatgcttg tgtgtcactg gtgttctcct
     4621 tcctctacaa ggtggtacag gtcttctccg agtactttaa ggagttggag gaggagagca
     4681 tccgagacaa ctttgtcatc atctacgagc tgctagatga actcatggac tttggctacc
     4741 cgcagaccac tgacagcaag atcttgcagg agtacatcac tcaggaaggc cacaagctgg
     4801 aaacgggggc ccctaggccc ccagccacag tcaccaatgc tgtgtcctgg cgttcagaag
     4861 gcatcaagta tcggaagaat gaagtattcc tggatgtcat tgaggctgtt aacctcttgg
     4921 tcagtgccaa tggcaacgtg ctgcgcagtg agattgtggg ttccatcaag atgcgggtct
     4981 tcctctcagg catgcctgag ttacgcctgg gtctcaatga caaggtcctc ttcgacaaca
     5041 caggccgagg gaagagcaag tcagtggagc tggaggatgt gaaattccac cagtgtgtgc
     5101 ggctgtcacg ttttgagaac gaccgcacta tctccttcat cccacccgac ggagagtttg
     5161 aactcatgtc ctaccgcctc aacacccatg tgaagccttt gatctggatt gagtccgtga
     5221 ttgagaagca ttcccacagc cgcattgagt acatggtcaa ggccaagagc cagttcaaga
     5281 ggcggtcaac agccaacaat gtagagatcc atataccagt ccccaacgat gctgattcac
     5341 ccaagttcaa gactacagtg gggagtgtca agtgggtccc tgaaaacagt gagatcgtgt
     5401 ggtccgtcaa gtcctttccg ggtggcaagg agtacctgat gcgggcccac tttggccttc
     5461 ccagtgtgga agctgaagac aaggagggaa agccccccat cagcgtcaag tttgagatcc
     5521 cctacttcac tacctctggc atccaggtgc gctacctgaa aatcattgag aagagtgggt
     5581 accaggccct gccctgggta cgatatatca cacagaacgg agattatcag ctccggaccc
     5641 agtgaacgcg tctcgagaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     5701 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     5761 ggttttttgc tgaaaggagg aactatatcc gactagtagc ccgcctaatg agcgggcttt
     5821 tttttaattc ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5881 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5941 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6001 cagaaacgct cgtgaaagta aaagacgcag aggaccaatt gggggcacga gtgggataca
     6061 tagaactgga cttgaatagc ggtaaaatcc ttgagagttt tcgccctgaa gagcgttttc
     6121 caatgatgag cactttcaaa gttctgctat gtggagcagt attatcccgt gtagatgcgg
     6181 ggcaagagca actcggacga cgaatacact attcgcagaa tgacttggtt gaatactccc
     6241 cagtgacaga aaagcacctt acggacggaa tgacggtaag agaattatgt agtgccgcca
     6301 taacgatgag tgataacact gcggcgaact tacttctgac aaccatcggt ggaccgaagg
     6361 aattaaccgc ttttttgcac aatatgggag accatgtaac tcgccttgac cgttgggaac
     6421 cagaactgaa tgaagccata ccaaacgacg agcgagacac cacaatgcct gcggcaatgg
     6481 caacaacatt acgcaaacta ttaactggcg aactacttac tctggcttca cggcaacaat
     6541 taatagactg gcttgaagcg gataaagttg caggaccact actgcgttcg gcacttcctg
     6601 ctggctggtt tattgctgat aaatctgggg caggagagcg tggttcacgg ggtatcattg
     6661 ccgcacttgg accagatggt aagccttccc gtatcgtagt tatctacacg acgggtagtc
     6721 aggcaactat ggacgaacga aatagacaga ttgctgaaat aggggcttca ctgattaagc
     6781 attggtaaac cgatacaatt aaaggctcct tttggagcct ttttttttgg acggaccggt
     6841 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6901 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6961 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     7021 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7081 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7141 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7201 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7261 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7321 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7381 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7441 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7501 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7561 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7621 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     7681 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     7741 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     7801 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     7861 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     7921 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     7981 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     8041 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     8101 tggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     8161 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     8221 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttgg
     8281 caccatgcat cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc
     8341 cctctagaaa taattttgtt taactttaag aaggagatat acatatgata catttcatat
     8401 tgctcttcag tcgacaaggg aaattacggc tacagaaatg gtacatcact ctccctgata
     8461 aagagaggaa gaagatcacc cgggaaattg ttcagattat tctctcccgt ggtcacagga
     8521 caagcagttt tgttgactgg aaggagctaa aacttgttta taaaaggtat gctagtttat
     8581 atttttgctg tgcaatagaa aatcaggaca atgaactctt gacgctagag attgtgcatc
     8641 gttacgtgga gctgctggac aaatattttg gaaatgtctg tgagctggat attatcttta
     8701 attttgaaaa ggcttatttc atcctggacg agtttataat aggtggggaa attcaggaaa
     8761 catccaagaa aattgctgtc aaagccattg aagactctga tatgttacag gaggtcagta
     8821 cggtttgcca aaccatggga gaaagatgaa gatcttgtaa aacgacggcc agtgaattca
     8881 ataattttgt ttaactttaa gaaggagata tacatatggg cagcagccat catcatcatc
     8941 atcacagcag cggatctggt ggtggtggtg gtgaaaacct gtacttccag g
//
