LOCUS       sequence-342026-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  VHS-GAT domain in GGA3 can bind to an active form of ARF1 protein. 
            Using this construct, you can assess the activity of ARF1 protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  AUTHORS   Dell'Angelica EC, Puertollano R, Mullins C, Aguilar RC, Vargas JD, 
            Hartnell LM, Bonifacino JS
  TITLE     GGAs: a family of ADP ribosylation factor-binding proteins related 
            to adaptors and associated with the Golgi complex.
  JOURNAL   J Cell Biol. 2000 Apr 3;149(1):81-94. doi: 10.1083/jcb.149.1.81.
  PUBMED    10747089
REFERENCE   2  (bases 1 to 5912)
  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: "10.1083/jcb.149.1";
            journalName: "J Cell Biol"; date: "2000-04-3- 3"; volume: "149"; 
            issue: "1"; pages: "81-94"
COMMENT     Sequence Label: GST-GGA3-VHS-GAT
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(670..675,676..693,694..700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    708..724
                     /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     732..748
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     732..748
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             744..1271
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(764..781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(764..780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(773..795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1373..1378,1379..1394,1395..1401)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1409..1425
                     /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             1448..2101
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2059..2081
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2111..2122
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3152..3174)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3319..3337)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3405..3509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3510..3578,3579..4370)
                     /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(3728..3747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4541..5129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5030..5049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5283..5300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5373..5450
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5451..5912,1..621)
                     /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(5470..5489)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
       61 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      121 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      181 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      241 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      301 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      361 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      421 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
      481 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
      541 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      601 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
      661 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      721 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
      781 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
      841 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
      901 gcagcctgaa tggcgaatgg cgctttgcct ggtttccggc accagaagcg gtgccggaaa
      961 gctggctgga gtgcgatctt cctgaggccg atactgtcgt cgtcccctca aactggcaga
     1021 tgcacggtta cgatgcgccc atctacacca acgtgaccta tcccattacg gtcaatccgc
     1081 cgtttgttcc cacggagaat ccgacgggtt gttactcgct cacatttaat gttgatgaaa
     1141 gctggctaca ggaaggccag acgcgaatta tttttgatgg cgttggaatt agcttatcga
     1201 ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt
     1261 gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat
     1321 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat
     1381 taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     1441 agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg
     1501 acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg
     1561 tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta
     1621 tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa
     1681 gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc
     1741 ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct
     1801 caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg
     1861 tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc
     1921 tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg
     1981 ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta
     2041 tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa
     2101 atcggatctg atcgaaggtc gtgggatccc cgaattctgc aatatggcgg aggcggaagg
     2161 ggaaagcctg gagtcctggc tcaataaagc caccaatcct tccaaccgcc aggaggactg
     2221 ggaatacata attggcttct gtgatcagat caacaaggag ctggaagggc cacagatcgc
     2281 cgtccgactg ctggcccaca agatccagtc cccacaggaa tgggaggcgc tccaggccct
     2341 gacggtgctg gaggcatgca tgaagaactg tgggaggaga tttcataacg aagtggggaa
     2401 gttccgcttt ttgaatgagt taatcaaagt cgtctctcca aagtacctgg gggacagggt
     2461 gtctgagaaa gtgaagacca aggttattga gctgctgtac agctggacca tggccctgcc
     2521 agaagaagca aagatcaaag acgcctacca catgctgaag agacagggca tagtgcagtc
     2581 tgacccacca attcctgtgg ataggacgct gatcccctct ccaccacctc gtcccaaaaa
     2641 ccctgttttt gatgatgagg agaagtccaa gcttttagcc aagctgctga aaagcaaaaa
     2701 cccagatgac ctgcaggagg ccaacaagct catcaagtcc atggtgaagg aagacgaggc
     2761 acggatccag aaggtgacca agcgtctgca cacgttagag gaagttaaca acaacgtgag
     2821 actgctcagt gagatgctgc ttcattacag ccaggaggac tcttcggacg gggacagaga
     2881 gctgatgaag gagctgtttg atcagtgtga gaacaagagg cggactttat ttaaactcgc
     2941 cagtgagact gaggacaatg ataacagttt gggggacatc ctgcaagcca gtgacaacct
     3001 ctcccgggtc atcaactctt acaaaacaat tattgaaggg caggtcatca atggcgaggt
     3061 ggctacctta accctgcctg actgagtcga cgcggccgca tcgtgactga ctgacgatct
     3121 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     3181 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     3241 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     3301 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     3361 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3481 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3601 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3661 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3721 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     3781 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3901 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4021 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     4081 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4201 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4381 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4441 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4501 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4561 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4621 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4681 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4741 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4801 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4861 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4921 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4981 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5041 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5101 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5161 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5221 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5281 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     5341 atctgtgcgg tatttcacac cgcataaatt ccgacaccat cgaatggcgc aaaacctttc
     5401 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     5461 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     5521 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     5581 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     5641 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     5701 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     5761 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     5821 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     5881 cggcgttatt tcttgatgtc tctgaccaga ca
//
