LOCUS       sequence-176206-        5508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GST-tagged GAT domain of GGA3 (107-286 aa) in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5508)
  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 5508)
  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/hGGA3(GAT)
FEATURES             Location/Qualifiers
     source          1..5508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     356..375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     609..626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        699..776
                     /gene="lacI"
                     /label=lacI promoter
     CDS             777..1859
                     /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(796..815)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1908..1913,1914..1931,1932..1938)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1946..1962
                     /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     1970..1986
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1970..1986
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1982..2509
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2002..2019)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2002..2018)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2033)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2611..2616,2617..2632,2633..2639)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2647..2663
                     /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             2686..3339
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3297..3319
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3346..3363
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3986..4008)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4153..4171)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4239..4343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4344..4412,4413..5204)
                     /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(4562..4581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5375..5508,1..455)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
       61 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      481 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      541 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      601 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
      661 ttacgcatct gtgcggtatt tcacaccgca taaattccga caccatcgaa tggcgcaaaa
      721 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
      781 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
      841 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
      901 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
      961 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     1021 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     1081 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     1141 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     1201 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     1261 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     1321 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     1381 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     1441 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     1501 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     1561 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     1621 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     1681 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     1741 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     1801 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     1861 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cccaggcttt acactttatg
     1921 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     1981 tatgaccatg attacggatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2041 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2101 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     2161 ctttgcctgg tttccggcac cagaagcggt gccggaaagc tggctggagt gcgatcttcc
     2221 tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg cacggttacg atgcgcccat
     2281 ctacaccaac gtgacctatc ccattacggt caatccgccg tttgttccca cggagaatcc
     2341 gacgggttgt tactcgctca catttaatgt tgatgaaagc tggctacagg aaggccagac
     2401 gcgaattatt tttgatggcg ttggaattag cttatcgact gcacggtgca ccaatgcttc
     2461 tggcgtcagg cagccatcgg aagctgtggt atggctgtgc aggtcgtaaa tcactgcata
     2521 attcgtgtcg ctcaaggcgc actcccgttc tggataatgt tttttgcgcc gacatcataa
     2581 cggttctggc aaatattctg aaatgagctg ttgacaatta atcatcggct cgtataatgt
     2641 gtggaattgt gagcggataa caatttcaca caggaaacag tattcatgtc ccctatacta
     2701 ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa
     2761 gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag
     2821 tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta
     2881 acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt
     2941 ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt
     3001 gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag
     3061 ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt
     3121 gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg
     3181 gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct
     3241 atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc
     3301 tggcaagcca cgtttggtgg tggcgaccat cctccaaaat cggatctggt tccgcgtgga
     3361 tccagacagg gcatagtgca gtctgaccca ccaattcctg tggataggac gctgatcccc
     3421 tctccaccac ctcgtcccaa aaaccctgtt tttgatgatg aggagaagtc caagctttta
     3481 gccaagctgc tgaaaagcaa aaacccagat gacctgcagg aggccaacaa gctcatcaag
     3541 tccatggtga aggaagacga ggcacggatc cagaaggtga ccaagcgtct gcacacgtta
     3601 gaggaagtta acaacaacgt gagactgctc agtgagatgc tgcttcatta cagccaggag
     3661 gactcttcgg acggggacag agagctgatg aaggagctgt ttgatcagtg tgagaacaag
     3721 aggcggactt tatttaaact cgccagtgag actgaggaca atgataacag tttgggggac
     3781 atcctgcaag ccagtgacaa cctctcccgg gtcatcaact cttacaaaac aattattgaa
     3841 gggcaggtca tcaatggcga ggtggctacc ttaaccctgc ctgactcgga aggaaacagt
     3901 cagtagaatt cccgggtcga ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg
     3961 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     4021 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4081 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     4141 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4201 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4261 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4321 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4381 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4441 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4501 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4561 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     4621 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4681 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4741 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4801 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4861 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     4921 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4981 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5041 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5101 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5161 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5221 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5281 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5341 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5401 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5461 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcaga
//
