pGEX-6p-1-Caprin1-EGFP

LOCUS       pGEX-6p-1-Caprin1-EGFP             7855 bp DNA     circular SYN 03-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7855)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Apr 17, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             951..971
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             978..3104
                     /codon_start=1
                     /label=CAPRIN1 Codon
                     /translation="MPSATSHSGSGSKSSGPPPPSGSSGSEAAAGAGAAAPASQHPATG
                     TGAVQTEAMKQILGVIDKKLRNLEKKKGKLDDYQERMNKGERLNQDQLDAVSKYQEVTN
                     NLEFAKELQRSFMALSQDIQKTIKKTARREQLMREEAEQKRLKTVLELQYVLDKLGDDE
                     VRTDLKQGLNGVPILSEEELSLLDEFYKLVDPERDMSLRLNEQYEHASIHLWDLLEGKE
                     KPVCGTTYKVLKEIVERVFQSNYFDSTHNHQNGLCEEEEAASAPAVEDQVPEAEPEPAE
                     EYTEQSEVESTEYVNRQFMAETQFTSGEKEQVDEWTVETVEVVNSLQQQPQAASPSVPE
                     PHSLTPVAQADPLVRRQRVQDLMAQMQGPYNFIQDSMLDFENQTLDPAIVSAQPMNPTQ
                     NMDMPQLVCPPVHSESRLAQPNQVPVQPEATQVPLVSSTSEGYTASQPLYQPSHATEQR
                     PQKEPIDQIQATISLNTDQTTASSSLPAASQPQVFQAGTSKPLHSSGINVNAAPFQSMQ
                     TVFNMNAPVPPVNEPETLKQQNQYQASYNQSFSSQPHQVEQTELQQEQLQTVVGTYHGS
                     PDQSHQVTGNHQQPPQQNTGFPRSNQPYYNSRGVSRGGSRGARGLMNGYRGPANGFRGG
                     YDGYRPSFSNTPNSGYTQSQFSAPRDYSGYQRDGYQQNFKRGSGQSGPRGAPRGRGGPP
                     RPNRGMPQMNTQQVN"
     misc_feature    complement(3087..3125)
                     /label=Primer R for caprin1 in pGEX-6P-1
     misc_feature    3105..3143
                     /label=Primer F for mEGFP at N ter in GST-Tag
     CDS             3123..3839
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      3123..3126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    complement(3819..3857)
                     /label=Primer R for mEGFP at N ter in GST-Tag
     misc_feature    3837..3875
                     /label=Primer F for caprin1 in pGEX-6P-1
     promoter        4158..4262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4263..5123
                     /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"
     rep_origin      5294..5882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6126..6203
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6204..7286
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    7299..7320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7335..7365
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7373..7389
                     /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     7397..7413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             7409..7855
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     complement(7429..7445)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc gaaaacctgt
      961 atttccaggg cggtagtatg ccgagtgcca ccagccatag cggtagcggt agcaaaagta
     1021 gtggtccgcc gccgccgagc ggtagctcag gttcagaagc cgcagccggt gccggtgcag
     1081 ccgcacctgc aagtcagcat ccggcaaccg gtacaggtgc cgttcagacc gaagcaatga
     1141 aacagattct gggcgttatt gataaaaaac tgcgcaatct ggaaaagaaa aaaggtaaac
     1201 tggatgatta ccaggaacgc atgaataagg gcgaacgtct gaatcaggat cagctggatg
     1261 ccgtgagtaa atatcaggaa gttaccaata atctggaatt tgccaaagaa ctgcagcgta
     1321 gttttatggc cctgagccag gatattcaga aaaccattaa gaaaaccgca cgccgtgaac
     1381 agctgatgcg tgaagaagcc gaacagaaac gtctgaaaac cgttctggaa ctgcagtatg
     1441 tgctggataa actgggtgac gatgaagttc gtaccgatct gaaacagggt ctgaatggtg
     1501 tgccgattct gagtgaagaa gaactgagcc tgctggatga attttataaa ctggtggacc
     1561 ctgaacgtga tatgagcctg cgtctgaatg aacagtatga acatgccagc attcatctgt
     1621 gggatctgct ggaaggtaaa gaaaaaccgg tttgtggcac cacctataaa gttctgaaag
     1681 aaattgtgga acgtgttttt cagagtaatt attttgacag cacccataat catcagaatg
     1741 gcctgtgtga agaagaagaa gcagcaagtg ccccggccgt ggaagatcag gttccggaag
     1801 cagaaccgga accggccgaa gaatataccg aacagagcga agtggaaagt accgaatatg
     1861 tgaatcgcca gtttatggcc gaaacccagt ttaccagtgg cgaaaaagaa caggtggatg
     1921 aatggaccgt tgaaaccgtt gaagttgtga atagcctgca gcagcagccg caggcagcaa
     1981 gcccgagcgt gcctgaaccg catagtctga ccccggttgc acaggcagat ccgctggtgc
     2041 gccgccagcg tgttcaggat ctgatggcac agatgcaggg cccgtataat tttattcagg
     2101 atagcatgct ggattttgaa aatcagaccc tggaccctgc cattgttagc gcccagccga
     2161 tgaatccgac ccagaatatg gatatgccgc agctggtgtg cccgccggtg catagtgaaa
     2221 gtcgtctggc ccagccgaat caggttccgg ttcagccgga agcaacccag gtgccgctgg
     2281 ttagtagtac cagcgaaggt tataccgcaa gtcagccgct gtatcagccg agccatgcca
     2341 ccgaacagcg tccgcagaaa gaaccgattg atcagattca ggccaccatt agtctgaata
     2401 ccgatcagac caccgccagt agcagcctgc cggccgctag tcagccgcag gtttttcagg
     2461 caggcaccag caaaccgctg catagtagcg gtattaatgt taatgcagca ccgtttcaga
     2521 gtatgcagac cgtgtttaat atgaatgccc cggttccgcc ggtgaatgaa ccggaaaccc
     2581 tgaaacagca gaatcagtat caggccagct ataatcagag ttttagtagc cagccgcatc
     2641 aggttgaaca gaccgaactg cagcaggaac agctgcagac cgtggtgggc acctatcatg
     2701 gtagtccgga tcagagtcat caggtgaccg gcaatcatca gcagccgccg cagcagaata
     2761 ccggctttcc gcgcagcaat cagccgtatt ataatagccg tggcgtgagc cgtggtggta
     2821 gccgcggtgc acgtggtctg atgaatggtt atcgtggccc ggcaaatggc tttcgtggtg
     2881 gttatgatgg ctatcgtccg agctttagta ataccccgaa tagtggttat acccagagcc
     2941 agtttagcgc accgcgtgat tatagcggct atcagcgcga tggttatcag cagaatttta
     3001 aacgcggcag cggtcagagc ggtccgcgcg gtgcacctcg tggtcgtggt ggtccgccgc
     3061 gtcctaatcg cggcatgccg cagatgaata cccagcaggt gaatggtggt agtggtggta
     3121 gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3181 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     3241 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     3301 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     3361 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     3421 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3481 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3541 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3601 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3661 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3721 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     3781 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagc
     3841 tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg
     3901 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     3961 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     4021 tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga
     4081 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4141 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4201 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4261 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4321 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4381 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4441 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4501 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4561 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4621 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4681 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4741 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4801 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4861 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4921 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4981 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5041 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5101 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5161 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5221 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5401 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5461 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5761 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5821 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5881 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5941 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6001 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6061 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
     6121 attccgacac catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     6181 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     6241 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     6301 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     6361 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     6421 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     6481 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     6541 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     6601 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     6661 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     6721 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     6781 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     6841 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     6901 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     6961 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     7021 acgacgatac cgaagacagc tcatgttata tcccgccgtc aaccaccatc aaacaggatt
     7081 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     7141 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     7201 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7261 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7321 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7381 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     7441 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     7501 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     7561 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     7621 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     7681 gcagatgcac ggttacgatg cgcccatcta caccaacgta acctatccca ttacggtcaa
     7741 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     7801 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaatt
//