LOCUS       sequence-387045-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-AP-4 beta-4 hinge-ear (residues 534-739) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  AUTHORS   Mattera R, Guardia CM, Sidhu SS, Bonifacino JS
  TITLE     Bivalent Motif-Ear Interactions Mediate the Association of the 
            Accessory Protein Tepsin with the AP-4 Adaptor Complex.
  JOURNAL   J Biol Chem. 2015 Dec 25;290(52):30736-49. doi: 
            10.1074/jbc.M115.683409. Epub 2015 Nov 5.
  PUBMED    26542808
REFERENCE   2  (bases 1 to 5608)
  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: 
            "10.1074/jbc.M115.683409"; journalName: "J Biol Chem"; date: 
            "2015-12-25- 25"; volume: "290"; issue: "52"; pages: "30736-49"
COMMENT     Sequence Label: pGEX-4T-1-beta-4 hinge-ear
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(182..187,188..203,204..210)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    218..234
                     /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)."
     CDS             257..910
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     868..890
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             917..934
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1657..1679)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1824..1842)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1910..2014
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2015..2083,2084..2875)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2233..2252)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3046..3634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3535..3554
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3788..3805
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3878..3955
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3956..5038
                     /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     complement(3975..3994)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5087..5092,5093..5110,5111..5117)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5125..5141
                     /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     5149..5165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5149..5165
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5161..5608,1..80)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5181..5198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5181..5197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5190..5212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
       61 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
      121 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
      181 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
      241 acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca
      301 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
      361 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
      421 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
      481 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
      541 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
      601 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
      661 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
      721 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
      781 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
      841 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
      901 tcctccaaaa tcggatctgg ttccgcgtgg atccccggaa ttccctatgg ccatggaggc
      961 ctgtagccct aaatctgacc ctactcttgg acttttggag gatccggcag aaagacctgt
     1021 gaatagctgg gcctcagact tcaacacact ggtgccagtg tatggcaaag cccactgggc
     1081 aactatctct aaatgccagg gggcagagcg ttgtgaccca gagcttccta aaacttcatc
     1141 ctttgccgca tcaggaccct tgattcctga agagaacaag gagagggtac aagaactccc
     1201 tgattctgga gccctcatgc tagtccccaa tcgccagctt actgctgatt attttgagaa
     1261 aacttggctt agccttaaag ttgctcatca gcaagtgttg ccttggcggg gagaattcca
     1321 tcctgacacc ctccagatgg ctcttcaagt agtgaacatc cagaccatcg caatgagtag
     1381 ggctgggtct cggccatgga aagcatacct cagtgctcag gatgatactg gctgtctgtt
     1441 cttaacagaa ctgctattgg agcctggaaa ctcagaaatg cagatctctg tgaaacaaaa
     1501 tgaagcaaga acggagacgc tgaatagttt tatttctgta ttagaaactg tgattggaac
     1561 aattgaagaa ataaaatcat aaccgggtcg actcgagcgg ccgcatcgtg actgactgac
     1621 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1681 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1741 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     1801 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     1861 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1921 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1981 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2041 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2101 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2161 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2221 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     2281 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2341 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2401 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2461 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2521 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     2581 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2641 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2701 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2761 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2821 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2881 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2941 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3001 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3061 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3121 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3181 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3601 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3661 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3721 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3781 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     3841 tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat ggcgcaaaac
     3901 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     3961 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4021 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4081 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4141 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4201 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4261 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4321 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4381 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     4441 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     4501 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     4561 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     4621 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     4681 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     4741 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     4801 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     4861 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     4921 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     4981 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5041 gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5101 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5161 atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5221 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5281 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     5341 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     5401 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     5461 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     5521 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     5581 cgaattattt ttgatggcgt tggaatta
//
