LOCUS       sequence-339777-        8296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing c-terminal V5-tagged GPR3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8296)
  AUTHORS   Iorio C, Rourke JL, Wells L, Sakamaki JI, Moon E, Hu Q, Kin T, 
            Screaton RA
  TITLE     Silencing the G-protein coupled receptor 3-salt inducible kinase 2 
            pathway promotes human beta cell proliferation.
  JOURNAL   Commun Biol. 2021 Jul 23;4(1):907. doi: 10.1038/s42003-021-02433-2.
  PUBMED    34302056
REFERENCE   2  (bases 1 to 8296)
  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: "Commun 
            Biol."; date: "2021-07-23"; volume: "4(1):907. doi"; pages: " 
            10.1038/s42003-021-02433-2"
COMMENT     Sequence Label: pLenti6.3delta4 GPR3-V5
FEATURES             Location/Qualifiers
     source          1..8296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(277..865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(357..376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1036..1827,1828..1896))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1659..1678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1897..2001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2027..2482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2164..2185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2376..2395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2609..2631
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2623..2640
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2624..2640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2650..2669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2650..2668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2689..2824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2864..2985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2875..2894)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2929..2948
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3057..3290
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     CDS             complement(3377..3775)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(3794..3841)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3889..4205)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3903..4038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4220..4808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4326..4337
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4735..4755
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4832..4873)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    4926..4950
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5944..5968)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6064..6088)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(6085..6136)
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(6114..6134)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    6186..6303
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6570..6614)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6799..7032
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7525..7650
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7697..7877
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7878..8104)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(8132..8152)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8132..8150)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8171..8187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8171..8187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8184..8206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8195..8211
                     /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)."
     promoter        complement(join(8219..8225,8226..8243,8244..8249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8264..8285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
       61 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
      901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     1741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     1801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     2041 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2101 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2161 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2221 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2281 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2341 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2401 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2461 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2521 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     2581 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2641 gagcgcgcgt aatacgactc actatagggc gaattgggta cgtccctagg cctccaaaaa
     2701 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     2761 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     2821 ggatgggcgg agttaggggc gggatagcta gagccagaca tgataagata cattgatgag
     2881 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2941 gctattgctt tatttgtaac cattataagc tgcaataaac aagttcctct cactctctga
     3001 tattcatttc tttgcaagtt ataaatactg aataataaga tgacatgaac tactactgct
     3061 agagattttc cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa
     3121 cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg
     3181 gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt
     3241 acaagcaaaa agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt
     3301 ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaaggtac
     3361 cgagctcgaa ttgtgcttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     3421 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     3481 cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     3541 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     3601 gttctgcaca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     3661 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     3721 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggttt
     3781 agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa
     3841 cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc
     3901 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc
     3961 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     4021 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     4081 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     4141 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     4201 acaccggtac catcgccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg
     4261 cgaaggcgaa gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc
     4321 gctggattga gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca
     4381 acacaggcga gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca
     4441 gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc
     4501 gtggcaatag ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg
     4561 ccccaaccag tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca
     4621 acgggccaca actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg
     4681 aaagccatac gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc
     4741 gtaaaaggag caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga
     4801 ggttgattgt cgatcattac taaccggtac gcgtagaatc gagaccgagg agagggttag
     4861 ggataggctt accttcgaac cgcgggccct ctagactcga gcggccgcca ctgtgctgga
     4921 tatcaaccac tttgtacaag aaagctgggt cgacatcact gggggagcgg gatcggaagg
     4981 ggatcttgga agaggaacag cagcagcaga cagcccacag cactttctgc acatcctggt
     5041 tgcggaaggc gtagatgata gggttgatca tggagttgta ggtggcaggg agcaaggtaa
     5101 gataggtgta gagaggtgga gagtgggcat cacccagcag gcagtagaca gtgaagggca
     5161 accagcaggc ggcaaaggct ccaagcacca cggccagtgt ggcaatgccc ttgcgggtgg
     5221 ccacatagtg ggaggcaggc agcaggtgcc gctgaagggc aatctgctgg gcatggcggc
     5281 agacgatgcg gcagatttgg gcgtagagct gcagcatgat gccaaacacc atgaagaagg
     5341 caatggccag aactaccaga tggttcttgg agagtggata aaccacgcca catgtggtca
     5401 ggccatccag gcagttccag gccagcacag gcagcagccc caggcccagg gcacctcccc
     5461 acactaaggc cagcatcaca taggtccgtg tcactgttgt ctctgaatag taggtgaggg
     5521 cgttatatag ggagagatac cggtcgacag tgatggccag tagactgccg atgctggcgg
     5581 taaaggccat tgccagcacg ccaaccagca ccaggctcat ctccgctgag ccgatgcaga
     5641 agacagcagc aaagtgcagg accaggccca ggcctgccag caggtctgcc acggccaggc
     5701 tgcccaccag caggaacatg ggggcacgga aggcaggagt gcccacgatg atggccacca
     5761 ctagcgcatt ctcgcaggac accagggtgc ctgagatgca gagcaccaca tcccaggcct
     5821 taggcgaggg cagtggtgcg gctggacctg tgggcccctc tgctgggccc acgctgctta
     5881 cattcacgtt gcctgagcca gctgagagcc aggccagagg gctgcctgca ccccacatca
     5941 tgaagcctgc ttttttgtac aaacttgttg atatctgcag aattccacca cactggacta
     6001 gtggatcctc tagagtcggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
     6061 ggcgatctga cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct
     6121 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgata
     6181 tcgataaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
     6241 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
     6301 aaagttaacc gataccgtcg agatccgttc actaatcgaa tggatctgtc tctgtctctc
     6361 tctccacctt cttcttctat tccttcgggc ctgtcgggtc ccctcggggt tgggaggtgg
     6421 gtctgaaacg ataatggtga atatccctgc ctaactctat tcactataga aagtacagca
     6481 aaaactattc ttaaacctac caagcctcct actatcatta tgaataattt tatataccac
     6541 agccaatttg ttatgttaaa ccaattccac aaacttgccc atttatctaa ttccaataat
     6601 tcttgttcat tcttttcttg ctggttttgc gattcttcaa ttaaggagtg tattaagctt
     6661 gtgtaattgt taatttctct gtcccactcc atccaggtcg tgtgattcca aatctgttcc
     6721 agagatttat tactccaact agcattccaa ggcacagcag tggtgcaaat gagttttcca
     6781 gagcaacccc aaatccccag gagctgttga tcctttaggt atctttccac agccaggatt
     6841 cttgcctgga gctgcttgat gccccagact gtgagttgca acagatgctg ttgcgcctca
     6901 atagccctca gcaaattgtt ctgctgctgc actataccag acaataattg tctggcctgt
     6961 accgtcagcg tcattgacgc tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg
     7021 aacaaagctc ctattcccac tgctcttttt tctctctgca ccactcttct ctttgccttg
     7081 gtgggtgcta ctcctaatgg ttcaattttt actactttat atttatataa ttcacttctc
     7141 caattgtccc tcatatctcc tcctccaggt ctgaagatca gcggccgctt gctgtgcggt
     7201 ggtcttactt ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat
     7261 ctctatcctt tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc
     7321 ttctgatcct gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga
     7381 tgtttctaac aggccaggat taactgcgaa tcgttctagc tccctgcttg cccatactat
     7441 atgttttaat ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga
     7501 tctaattctc ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc
     7561 gctagtcaaa atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc
     7621 ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc
     7681 cctgttcggg cgccactgct agagattttc cacactgact aaaagggtct gagggatctc
     7741 tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt
     7801 tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg
     7861 tctaaccaga gagacccgtt tatgtatcga gctaggcact taaatacaat atctctgcaa
     7921 tgcggcaatt cagtggttcg tccaatccat gtcagacccg tctgttgcct tcctaataag
     7981 gcacgatcgt accaccttac ttccaccaat cggcatgcac ggtgcttttt ctctccttgt
     8041 aaggcatgtt gctaactcat cgttaccatg ttgcaagact acaagagtat tgcataagac
     8101 tacattaagc ttgcagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     8161 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8221 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8281 cattaattgc gttgcg
//
