LOCUS       sequence-176200-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-tagged GAT domain of GGA1 (141-327 aa) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  AUTHORS   Takatsu H, Yoshino K, Toda K, Nakayama K
  TITLE     GGA proteins associate with Golgi membranes through interaction 
            between their GGAH domains and ADP-ribosylation factors.
  JOURNAL   Biochem J. 2002 Jul 15;365(Pt 2):369-78.
  PUBMED    11950392
REFERENCE   2  (bases 1 to 5528)
  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: "Biochem 
            J."; date: "2002-07-15"; volume: "365(Pt 2)"; pages: "369-78"
COMMENT     Sequence Label: pGEX-4T-2/hGGA1(GAT)
FEATURES             Location/Qualifiers
     source          1..5528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      578..1166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1067..1086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1320..1337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1410..1487
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1488..2570
                     /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(1507..1526)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2619..2624,2625..2642,2643..2649)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2657..2673
                     /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     2681..2697
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2681..2697
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2693..3220
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2713..2730)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2713..2729)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2744)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(3322..3327,3328..3343,3344..3350)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3358..3374
                     /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             3397..4050
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     4008..4030
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             4057..4074
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4717..4739)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4884..4902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4970..5074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5075..5143,5144..5528,1..407)
                     /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(5293..5312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
       61 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
      121 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      181 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      241 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      301 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      361 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
      421 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
      481 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
      541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
      661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
      721 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
      781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
      841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
      901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
      961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1261 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1321 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     1381 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg
     1441 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     1501 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     1561 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     1621 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     1681 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     1741 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     1801 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     1861 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     1921 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     1981 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     2041 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     2101 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     2161 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     2221 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     2281 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     2341 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     2401 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     2461 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2521 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2581 attaatgtaa gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2641 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     2701 gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     2761 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     2821 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg
     2881 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
     2941 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
     3001 cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
     3061 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
     3121 ttttgatggc gttggaatta gcttatcgac tgcacggtgc accaatgctt ctggcgtcag
     3181 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc
     3241 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     3301 caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg
     3361 tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact aggttattgg
     3421 aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat
     3481 gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg
     3541 ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct
     3601 atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag
     3661 cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga
     3721 attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa
     3781 atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta
     3841 acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg
     3901 tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa
     3961 attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc
     4021 acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg atccaagcag
     4081 gggattgtaa agtccgaccc caagcttcca gatgacacta cctttcccct tcctcctcca
     4141 cggccgaaga atgtgatctt tgaagatgag gagaaatcca agatgctggc ccgcctgctg
     4201 aagagctccc atcccgaaga cctccgcgca gccaataagc tcatcaaaga gatggtgcag
     4261 gaggaccaga agcggatgga gaagatctcg aagagggtga atgccatcga ggaggtgaac
     4321 aacaatgtga aactgctcac ggagatggtg atgagccaca gccagggcgg cgcagcagct
     4381 ggcagcagcg aggacctcat gaaggaactg taccagcgct gtgagcggat gcggcccacg
     4441 ctcttccgac tggcgagtga cacagaggac aatgatgagg ccttagcgga gatcctgcag
     4501 gccaatgaca acctcaccca ggtgatcaac ctgtataagc agctggtgcg gggtgaggag
     4561 gtcaacggtg atgccacagc cggctccatc cctgggagca cctcggccct gctggatctc
     4621 tcaggcctgg atctctgaat tcccgggtcg actcgagcgg ccgcatcgtg actgactgac
     4681 gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4741 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4801 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     4861 gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4921 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5041 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5161 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5281 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     5341 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5461 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5521 tttgcaca
//
