LOCUS       sequence-255129-        7654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon vector that Inducibly (Tet-On) expresses RTP1S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7654)
  AUTHORS   Jones EM, Jajoo R, Cancilla D, Lubock NB, Wang J, Satyadi M, Cheung 
            R, de March C, Bloom JS, Matsunami H, Kosuri S
  TITLE     A Scalable, Multiplexed Assay for Decoding GPCR-Ligand Interactions 
            with RNA Sequencing.
  JOURNAL   Cell Syst. 2019 Mar 27;8(3):254-260.e6. doi: 
            10.1016/j.cels.2019.02.009. Epub 2019 Mar 20.
  PUBMED    30904378
REFERENCE   2  (bases 1 to 7654)
  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.cels.2019.02.009"; journalName: "Cell Syst"; date: 
            "2019-03-27- 27"; volume: "8"; issue: "3"; pages: "254-260.e6"
COMMENT     Sequence Label: pB-RTP1S-Integration
FEATURES             Location/Qualifiers
     source          1..7654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    273..291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    347..365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    384..402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    421..439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    458..476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    495..513
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    532..550
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     572..596
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    1606..1740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1851..2062
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2075..2343
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2370..2969
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2370..2389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2866..2886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3029..3049)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3091..3212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3975..4009
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4500..4516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4500..4516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4513..4535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4524..4540
                     /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(4548..4554,4555..4572,4573..4578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4593..4614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4731..4748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4902..5490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4982..5001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5661..6452,6453..6521))
                     /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     6284..6303
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6522..6626)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6652..7107)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6789..6810)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7001..7020
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7234..7256
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7248..7265
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7249..7265
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   7395..7457
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 cccccaggga tgtaattacg tccctccccc gctagggggc agcagcgagc cgcccggggc
       61 tccgctccgg tccggcgctc cccccgcatc cccgagccgg cagcgtgcgg ggacagcccg
      121 ggcacgggga aggtggcacg ggatcgcttt cctctgaacg cttctcgctg ctctttgagc
      181 ctgcagacac ctggggggat acggggaaaa ggcctccacg gccactagtc caatcatagg
      241 ggtacccgac ccatcaagtg ccacctgacg tctccctatc agtgatagag aagtcgacac
      301 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
      361 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
      421 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
      481 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
      541 gtgatagaga aggtaccccc tatataagca gagctcgttt agtgaaccgt cagatcgcct
      601 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctga
      661 gctcgtttag tgaccgtcta gagccgccac catgtgtaag agtgtgacca caggtgagtg
      721 gaagaaggtc ttctacgaga agatggagga ggtgaagcca gcggacagct gggacttcat
      781 catagacccc aacctcaagc acaatgtgtt ggcccctggc tggaagcagt acctggaact
      841 tcatgcctca ggcaggttcc actgttcctg gtgctggcac acctggcagt caccccatgt
      901 agtcatcctc ttccacatgt acctggacaa ggctcagcgc gctggttcgg tgcgcatgcg
      961 tgtgttcaag cagctctgct acgagtgcgg tacagcacgg ctggatgagt ccagcatgct
     1021 ggaggagaac atcgaaagcc tggtggacaa cctcatcacc agtttgcgag agcagtgcta
     1081 cggggagcgt ggtggccact accgcatcca tgtggccagc cggcaggaca accggcgaca
     1141 ccgcggagag ttctgcgagg cctgccagga aggcatcgtg cactggaagc ccagtgagaa
     1201 gctgctggag gaggaggcga ccacctacac cttctcccgt gctcccagcc ccaccaaacc
     1261 gcaggctgaa acaggctcag gctgcaactt ctgctccatt ccctggtgct tattttgggc
     1321 cacggttttg atgctcatca tctacctgca attctccttc cgtacttctg tctgagaatt
     1381 cgccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     1441 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     1501 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     1561 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     1621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     1741 cccaggaagc tcctctgtgt cctcataaac cctaacctcc tctacttgag aggacattcc
     1801 aatcataggg cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg
     1861 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta
     1921 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     1981 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     2041 cgggtttgcc gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc
     2101 ccgccgccct acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc
     2161 ctgtggtgcc tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc
     2221 gggcctttgt ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg
     2281 cctgaccctg cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag
     2341 atccaagctg tgaccggcgc ctacgctaga tgaccgagta caagcccacg gtgcgcctcg
     2401 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     2461 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     2521 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     2581 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     2641 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     2701 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     2761 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     2821 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     2881 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     2941 ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat tacaaaattt
     3001 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3061 ctttaatgcc tttgtatcat gcgttaacta aacttgttta ttgcagctta taatggttac
     3121 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3181 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt
     3241 caattagtct atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta
     3301 atatttaaac agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg
     3361 ttgcagggtt tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt
     3421 tagccccgga attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta
     3481 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc
     3541 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg
     3601 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac
     3661 gtaattacat ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat
     3721 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     3781 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     3841 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     3901 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     3961 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct
     4021 gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta
     4081 tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa
     4141 agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat
     4201 gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac
     4261 ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg
     4321 catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa
     4381 tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc
     4441 gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg
     4501 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4561 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4621 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5341 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6661 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6721 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6781 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6841 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6901 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6961 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     7021 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     7081 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     7141 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     7201 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     7261 ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg
     7321 gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca
     7381 cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
     7441 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
     7501 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
     7561 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa
     7621 caaaactttt atgagggaca gccccccccc aaag
//
