LOCUS       sequence-431285-        7794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used to make a lenti virus for stable expression of phosducin-like 
            protein 1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7794)
  AUTHORS   Wang S, Sass MI, Kwon Y, Ludlam WG, Smith TM, Carter EJ, Gladden NE,
            Riggi M, Iwasa JH, Willardson BM, Shen PS
  TITLE     Visualizing the chaperone-mediated folding trajectory of the G 
            protein beta5 beta-propeller.
  JOURNAL   Mol Cell. 2023 Nov 2;83(21):3852-3868.e6. doi: 
            10.1016/j.molcel.2023.09.032. Epub 2023 Oct 17.
  PUBMED    37852256
REFERENCE   2  (bases 1 to 7794)
  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: 
            "10.1016/j.molcel.2023.09.032"; journalName: "Mol Cell"; date: 
            "2023-11-2- 2"; volume: "83"; issue: "21"; pages: "3852-3868.e6"
COMMENT     Sequence Label: pLenti-Blast PhLP1-myc-6xHis
FEATURES             Location/Qualifiers
     source          1..7794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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"
     promoter        2119..2322
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2273..2293
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2324..2342
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2345..2363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      2421..2430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3354..3383
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3399..3416
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3452..3469)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(3481..3501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3486..3815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3666..3801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3838..4236
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4315..4548
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4620..4741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4657..4676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4711..4730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4781..4916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4936..4955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4937..4955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4965..4982)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4965..4981)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4974..4996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5123..5578
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5210..5229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5420..5441
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5604..5708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5709..5777,5778..6569)
                     /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(5927..5946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6740..7328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7229..7248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7482..7499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7616..7637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7652..7657,7658..7675,7676..7682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7690..7706
                     /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     7695..7717
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7714..7730
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7714..7730
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7749..7769
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7751..7769
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccctttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgata
     1801 agcttgggag ttccgcttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1861 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1921 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1981 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     2041 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     2101 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     2161 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2221 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2281 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga
     2341 gatctcccta tcagtgatag agatcgtcga ctagtggatc cagtgtggtg gaattctgca
     2401 gatatccagc acagtggttc gccaccatgg gaaccaccct tgatgataag ttgctggggg
     2461 agaaactgca gtactactat agcagcagtg aggatgagga cagtgaccac gaggacaagg
     2521 accgaggcag atgtgcccca gccagcagtt ctgtgcctgc agaggctgag ctggcaggcg
     2581 aaggcatctc agttaacaca ggcccaaaag gtgtgatcaa tgactggcgc cgcttcaagc
     2641 agttggagac agagcagagg gaggagcagt gccgggagat ggaaaggctg atcaagaagc
     2701 tgtcaatgac ttgcaggtcc catctggatg aagaggagga gcaacagaaa cagaaagacc
     2761 tccaggagaa gatcagtggg aagatgactc tgaaggagtt tgccataatg aatgaggacc
     2821 aagatgatga agagtttctg cagcagtacc ggaagcagcg aatggaagag atgcggcagc
     2881 agcttcacaa ggggccccaa ttcaagcagg tttttgagat ctccagtgga gaagggtttt
     2941 tagacatgat tgataaagaa cagaaaagca ttgtcatcat ggttcatatt tatgaggatg
     3001 gcattccagg gaccgaagcc atgaatggtt gcatgatctg ccttgccgca gagtacccag
     3061 ctgtcaagtt ctgcaaggtg aagagctcag ttattggcgc cagcagtcag ttcaccagga
     3121 atgcccttcc tgccctgctg atctataagg ggggtgaatt gatcggcaat tttgttcgtg
     3181 ttactgacca gctgggggat gatttctttg ctgtggacct tgaagctttt ctccaggaat
     3241 ttggattact cccagaaaag gaagtcttgg tgctgacatc tgtgcgtaac tctgccacgt
     3301 gtcacagtga ggatagcgac ctggaaatag atgctctaga gggcccgcgg ttcgaacaaa
     3361 aactcatctc agaagaggat ctgaatatgc ataccggtca tcatcaccat caccattgag
     3421 tttaaacccg ctgattaaac ctgctgatca gcctcgactg tgccttctag tcgaccctgt
     3481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3661 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3721 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     3781 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taccgaatcc tggaccgatg
     3841 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     3901 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     3961 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     4021 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     4081 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     4141 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     4201 gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgggtacct ttaagaccaa
     4261 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4321 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     4381 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4441 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     4501 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     4561 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     4621 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4681 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4741 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     4801 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4861 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4921 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4981 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5041 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5101 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5161 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5221 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5281 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5341 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5401 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5461 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5521 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5701 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5761 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5821 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5881 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5941 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6001 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6061 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6121 caacgatcgg aggacctaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6181 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6241 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6301 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6361 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6421 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6481 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6541 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6601 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6661 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6721 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6781 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6841 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6901 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6961 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7021 gacgatagtt accggataac gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7081 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7141 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7201 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7261 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7321 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7381 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7441 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     7501 aagcggaaga gcgcccaata cgcaaaccgc ccctccccgc gcgttggccg attcattaat
     7561 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7621 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     7681 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     7741 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gctt
//
