LOCUS       sequence-387073-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-AP-4 epsilon ear (residues 840-1137) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  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 5853)
  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-epsilon ear
FEATURES             Location/Qualifiers
     source          1..5853
                     /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(1902..1924)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2069..2087)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2155..2259
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2260..2328,2329..3120)
                     /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(2478..2497)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3291..3879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3780..3799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4033..4050
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4123..4200
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4201..5283
                     /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(4220..4239)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5332..5337,5338..5355,5356..5362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5370..5386
                     /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     5394..5410
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5394..5410
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5406..5853,1..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(5426..5443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5426..5442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5435..5457)
                     /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 atccgacaag gaattaaaga aattttctct
      961 cacttcagaa cttttggatt ctgagtcact cacagaactg cccttggttg agaaattctc
     1021 atattgtagt ctgtctacac cttcattgtt tgctaataac aacatggaaa tttttcaccc
     1081 tcctcaatct actgcagcct cagttgccaa ggaaagctct ttagcttcat cctttttgga
     1141 agaaactact gaatacatac actcaaatgc tatggaagtc tgtaataatg aaactatatc
     1201 agtgtcttct tataaaattt ggaaagatga ttgtttattg atggtctggt cagtcactaa
     1261 taagagtggt ttggaattga aaagtgctga cttagaaatt tttcctgcag aaaatttcaa
     1321 ggtgactgag caacctggat gctgtttgcc tgtaatggaa gcagaaagca ccaaaagctt
     1381 tcaatatagt gtgcagatag aaaaaccttt tacagaagga aatcttactg gttttattag
     1441 ttatcatatg atggatactc attctgctca gctggaattt tctgtaaact tatcactatt
     1501 agatttcatt agaccattaa aaatctcaag tgacgacttt gggaaactct ggttatcctt
     1561 cgcaaatgat gtgaaacaaa atgtaaaaat gtcagaatct caagctgcac ttccttctgc
     1621 actaaagact ctgcaacaga aactaagact ccatattatt gagattatag gcaatgaagg
     1681 gctattggcc tgtcagctgc tcccatccat cccctgctta ctgcattgcc gagttcatgc
     1741 agatgtatta gccctgtggt tcagatcctc ctgttctact cttcctgact atttactgta
     1801 tcaatgtcaa aaggtgatgg agggatctta ggtcgactcg agcggccgca tcgtgactga
     1861 ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1921 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1981 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     2041 ggagtgtata attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     2101 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2161 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2221 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2281 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2341 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2401 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2461 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2521 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2581 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2641 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2701 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2761 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     2821 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2881 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2941 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3001 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3061 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3121 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3181 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3241 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3361 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3421 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3481 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3541 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3601 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3661 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3721 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3781 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3841 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3901 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3961 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4021 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4081 ctccttacgc atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc
     4141 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     4201 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     4261 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     4321 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     4381 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     4441 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     4501 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     4561 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     4621 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     4681 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     4741 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     4801 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     4861 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     4921 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     4981 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     5041 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     5101 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     5161 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     5221 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     5281 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt
     5341 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     5401 cagctatgac catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5461 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5521 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     5581 ggcgctttgc ctggtttccg gcaccagaag cggtgccgga aagctggctg gagtgcgatc
     5641 ttcctgaggc cgatactgtc gtcgtcccct caaactggca gatgcacggt tacgatgcgc
     5701 ccatctacac caacgtgacc tatcccatta cggtcaatcc gccgtttgtt cccacggaga
     5761 atccgacggg ttgttactcg ctcacattta atgttgatga aagctggcta caggaaggcc
     5821 agacgcgaat tatttttgat ggcgttggaa tta
//
