LOCUS       sequence-233636-        9597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPI reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9597)
  AUTHORS   Zhang Q, Zheng YW, Coughlin SR, Shu X
  TITLE     A rapid fluorogenic GPCR-beta-arrestin interaction assay.
  JOURNAL   Protein Sci. 2018 Apr;27(4):874-879. doi: 10.1002/pro.3385. Epub 
            2018 Feb 23.
  PUBMED    29411438
REFERENCE   2  (bases 1 to 9597)
  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: "10.1002/pro.3385"; 
            journalName: "Protein Sci"; date: "2018-04"; volume: "27"; issue: 
            "4"; pages: "874-879"
COMMENT     Sequence Label: pLenti-GFP1-9
FEATURES             Location/Qualifiers
     source          1..9597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    487..1075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(540..560)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(958..969)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1078..1094)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1079..1095)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1185..1418
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(1443..1461)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1444..1461)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1751..1770)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1984..2003)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2103..2125
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2163..2181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2249..2353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2354..2422,2423..3214)
                     /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     complement(2572..2591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3385..3973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3874..3893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4127..4144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4214..4234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4219..4548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4399..4534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          5682..5747
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5877..5897
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6322..6456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6359..6378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6413..6432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6624..7257
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7304..7429
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7926..8159
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8344..8388
                     /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    8673..8790
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8943..9350
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
ORIGIN
        1 gccgtggccg accctggtga ccaccctgac ctatggcgtg cagtgcttta gccgctatcc
       61 ggatcatatg aaacgccatg atttttttaa aagcgcgatg ccggaaggct atgtgcagga
      121 acgcaccatt tattttaaag atgatggcac ctataaaacc cgcgcggaag tgaaatttga
      181 aggcgatacc ctggtgaacc gcattgaact gaaaggcatt gattttaaag aagatggcaa
      241 cattctgggc cataaactgg aatataactt taacagccat aaagtgtata ttaccgcgga
      301 taaacagaac aacggcatta aagcgaactt taccattcgc cataacgtgg aagatggcag
      361 cgtgcagctg gcggatcatt atcagcagaa caccccgatt ggcgatggcc cggttcttct
      421 tccttgagcg gccgcgactc tagagtcgac ctgcaggcat gcaagcttga tatcaagctt
      481 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
      541 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
      601 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
      661 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
      721 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
      781 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
      841 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
      901 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
      961 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     1021 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     1081 taccgtcgac ctcgagggaa ttaattcgag ctcggtacct ttaagaccaa tgacttacaa
     1141 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     1201 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     1261 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     1321 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     1381 cctcagaccc ttttagtcag tgtggaaaat ctctagcagc atctagaatt aattccgtgt
     1441 attctatagt gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag
     1501 ccgcgttcta acgacaatat gtacaagcct aattgtgtag catctggctt actgaagcag
     1561 accctatcat ctctctcgta aactgccgtc agagtcggtt tggttggacg aaccttctga
     1621 gtttctggta acgccgtccc gcacccggaa atggtcagcg aaccaatcag cagggtcatc
     1681 gctagccaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
     1741 gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg
     1801 ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg
     1861 ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta
     1921 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaat ggtgcactct
     1981 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     2041 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2101 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     2161 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     2221 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2281 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2341 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2401 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2461 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2521 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2581 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2641 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2701 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2761 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2821 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2881 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2941 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3001 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3061 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     3181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     3241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     4081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     4141 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     4201 ttaatgcagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttggacac
     4561 aagacaggct tgcgagatat gtttgagaat accactttat cccgcgtcag ggagaggcag
     4621 tgcgtaaaaa gacgcggact catgtgaaat actggttttt agtgcgccag atctctataa
     4681 tctcgcgcaa cctattttcc cctcgaacac tttttaagcc gtagataaac aggctgggac
     4741 acttcacatg agcgaaaaat acatcgtcac ctgggacatg ttgcagatcc atgcacgtaa
     4801 actcgcaagc cgactgatgc cttctgaaca atggaaaggc attattgccg taagccgtgg
     4861 cggtctgtac cgggtgcgtt actggcgcgt gaactgggta ttcgtcatgt cgataccgtt
     4921 tgtatttcca gctacgatca cgacaaccag cgcgagctta aagtgctgaa acgcgcagaa
     4981 ggcgatggcg aaggcttcat cgttattgat gacctggtgg ataccggtgg tactgcggtt
     5041 gcgattcgtg aaatgtatcc aaaagcgcac tttgtcacca tcttcgcaaa accggctggt
     5101 cgtccgctgg ttgatgacta tgttgttgat atcccgcaag atacctggat tgaacagccg
     5161 tgggatatgg gcgtcgtatt cgtcccgcca atctccggtc gctaatcttt tcaacgcctg
     5221 gcactgccgg gcgttgttct ttttaacttc aggcgggtta caatagtttc cagtaagtat
     5281 tctggaggct gcatccatga cacaggcaaa cctgagcgaa accctgttca aaccccgctt
     5341 taaacatcct gaaacctcga cgctagtccg ccgctttaat cacggcgcac aaccgcctgt
     5401 gcagtcggcc cttgatggta aaaccatccc tcactggtat cgcatgatta accgtctgat
     5461 gtggatctgg cgcggcattg acccacgcga aatcctcgac gtccaggcac gtattgtgat
     5521 gagcgatgcc gaacgtaccg acgatgattt atacgatacg gtgattggct accgtggcgg
     5581 caactggatt tatgagtggg ccccggatct ttgtgaagga accttacttc tgtggtgtga
     5641 cataattgga caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa
     5701 gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt ccaacctatg
     5761 gaactgatga atgggagcag tggtggaatg cctttaatga ggaaaacctg ttttgctcag
     5821 aagaaatgcc atctagtgat gatgaggcta ctgctgactc tcaacattct actcctccaa
     5881 aaaagaagag aaaggtagaa gaccccaagg actttccttc agaattgcta agttttttga
     5941 gtcatgctgt gtttagtaat agaactcttg cttgctttgc tatttacacc acaaaggaaa
     6001 aagctgcact gctatacaag aaaattatgg aaaaatattc tgtaaccttt ataagtaggc
     6061 ataacagtta taatcataac atactgtttt ttcttactcc acacaggcat agagtgtctg
     6121 ctattaataa ctatgctcaa aaattgtgta cctttagctt tttaatttgt aaaggggtta
     6181 ataaggaata tttgatgtat agtgccttga ctagagatca taatcagcca taccacattt
     6241 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     6301 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     6361 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6421 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     6481 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     6541 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
     6601 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat
     6661 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     6721 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     6781 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     6841 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     6901 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     6961 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     7021 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     7081 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7141 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7201 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     7261 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     7321 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     7381 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     7441 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     7501 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     7561 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     7621 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     7681 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     7741 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     7801 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     7861 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     7921 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     7981 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     8041 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     8101 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     8161 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     8221 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     8281 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     8341 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     8401 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     8461 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     8521 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     8581 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     8641 cggtatcgcc aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     8701 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     8761 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     8821 gtttggatcg ataagcttga tatcgaattc ctgcagcccc gataaaataa aagattttat
     8881 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctg
     8941 cagtaacgcc attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat
     9001 caagggcggg tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag
     9061 tttcggcccc ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag
     9121 gccaagaaca gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca
     9181 gatgtttcca ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa
     9241 tcagcctgct tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc
     9301 acaacccctc actcggcgcg ccagtcctcc gacagactga gtcgcccggg ggggatctgg
     9361 agctctcgag aattctcacg cgtctgcagg atatcaagct tgcggtaccg cgggcccggg
     9421 atccgccgcc accatgcgca aaggcgaaga actgtttacc ggcattgtgc cgattctggt
     9481 ggaactggat ggcgatgtga acggccataa attttttgtg cgcggcgaag gcgaaggcga
     9541 tgcgaccatt ggcaaactga gcctgaaatt tatttgcacc accggcaaac tgccggt
//
