LOCUS       sequence-177985-        6015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A bacterial expression plasmid encoding retinitis pigmentosa 2 (RP2)
            with N-terminus GST-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6015)
  AUTHORS   Madugula V, Lu L
  TITLE     A ternary complex comprising transportin1, Rab8 and the ciliary 
            targeting signal directs proteins to ciliary membranes.
  JOURNAL   J Cell Sci. 2016 Oct 15;129(20):3922-3934. Epub 2016 Sep 15.
  PUBMED    27633000
REFERENCE   2  (bases 1 to 6015)
  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: "J Cell 
            Sci."; date: "2016-10-15"; volume: "129(20)"; pages: "3922-3934. 
            Epub 2016 Sep 15"
COMMENT     Sequence Label: GST-RP2
FEATURES             Location/Qualifiers
     source          1..6015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        840..917
                     /gene="lacI"
                     /label=lacI promoter
     CDS             918..2000
                     /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(937..956)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2049..2054,2055..2072,2073..2079)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2087..2103
                     /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     2111..2127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2111..2127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2123..2650
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2143..2160)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2143..2159)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2152..2174)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2752..2757,2758..2773,2774..2780)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2788..2804
                     /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             2827..3480
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3438..3460
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3487..3504
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4634..4656)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4801..4819)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4887..4991
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4992..5060,5061..5852)
                     /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(5210..5229)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
      841 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
      901 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
      961 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     1021 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     1081 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     1141 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     1201 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     1261 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     1321 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     1381 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     1441 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     1501 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     1561 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     1621 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     1681 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     1741 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     1801 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     1861 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     1921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1981 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     2041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     2101 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca
     2161 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2221 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2281 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag
     2341 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat
     2401 gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg tcaatccgcc
     2461 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag
     2521 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac
     2581 tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg
     2641 caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg
     2701 ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt
     2761 aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2821 gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga
     2881 cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt
     2941 gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat
     3001 attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag
     3061 cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg
     3121 gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc
     3181 aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt
     3241 cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct
     3301 cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt
     3361 tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat
     3421 atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa
     3481 tcggatctgg ttccgcgtgg atccccggaa ttcatgggct gcttcttctc caagagacgg
     3541 aaggctgaca aggagtcgcg gcccgagaac gaggaggagc ggccaaagca gtacagctgg
     3601 gatcagcgcg agaaggttga tccaaaagac tacatgttca gtggactgaa ggatgaaaca
     3661 gtaggtcgct tacctgggac ggtagcagga caacagtttc tcattcaaga ctgtgagaac
     3721 tgtaacatct atatttttga tcactctgct acagttacca ttgatgactg tactaactgc
     3781 ataatttttc tgggacccgt gaaaggcagc gtgtttttcc ggaattgcag agattgcaag
     3841 tgcacattag cctgccaaca atttcgtgtg cgagattgta gaaagctgga agtctttttg
     3901 tgttgtgcca ctcaacccat cattgagtct tcctcaaata tcaaatttgg atgttttcaa
     3961 tggtactatc ctgaattagc tttccagttc aaagatgcag ggctaagtat cttcaacaat
     4021 acatggagta acattcatga ctttacacct gtgtcaggag aactcaactg gagccttctt
     4081 ccagaagatg ctgtggttca ggactatgtt cctataccta ctaccgaaga gctcaaagct
     4141 gttcgtgttt ccacagaagc caatagaagc attgttccaa tatcccgggg tcagagacag
     4201 aagagcagcg atgaatcatg cttagtggta ttatttgctg gtgattacac tattgcaaat
     4261 gccagaaaac taattgatga gatggttggt aaaggctttt tcctagttca gacaaaggaa
     4321 gtgtccatga aagctgagga tgctcaaagg gtttttcggg aaaaagcacc tgacttcctt
     4381 cctcttctga acaaaggtcc tgttattgcc ttggagttta atggggatgg tgctgtagaa
     4441 gtatgtcaac ttattgtaaa cgagatattc aatgggacca agatgtttgt atctgaaagc
     4501 aaggagacgg catctggaga tgtagacagc ttctacaact ttgctgatat acagatggga
     4561 atagtcgact cgagcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg
     4621 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4681 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4741 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg
     4801 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4861 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4921 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5221 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5341 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5521 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5581 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5821 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5881 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6001 tagaaaagat caaag
//
