LOCUS       sequence-496938-        7671 bp    DNA     circular SYN 12-JAN-2026
DEFINITION  Expression of GPR161 wild type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Krantz S, Bell B, Lund K, Parra NS, Ng Y, De Oliveira Rosa N, 
            Mukhopadhyay S, Croix BS, Sarin KY, Weigert R, Raimondi F, 
            Iglesias-Bartolome R
  TITLE     Dissection of Galphas and Hedgehog signaling crosstalk reveals 
            therapeutic opportunities to target adenosine receptor 2b in 
            Hedgehog-dependent tumors.
  JOURNAL   bioRxiv [Preprint]. 2025 Feb 27:2025.02.21.639530. doi: 
            10.1101/2025.02.21.639530.
  PUBMED    40060632
REFERENCE   2  (bases 1 to 7671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 12, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2025 Feb 27:2025.02.21.639530. doi: 
            10.1101/2025.02.21.639530."
COMMENT     Sequence Label: pCEFL_GPR161-HA
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        298..1480
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          529..1471
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1428..1448
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3146..3172
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3224..3242)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3225..3242)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3262..3279)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3268..3492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3538..3966
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3625..3644)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3835..3856
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3975..3995)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3980..4309
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4160..4295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4222..4241
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4376..5170
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4430..4449)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5040..5059
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5344..5465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5381..5400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5435..5454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5514..5530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5514..5530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5527..5549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5538..5554
                     /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(5562..5568,5569..5586,5587..5592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5607..5628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5745..5762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5916..6504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5996..6015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6675..7466,7467..7535))
                     /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     7298..7317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7536..7640)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgagctttgc aaagatggat aaagttttaa
      241 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
      301 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      361 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      421 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      481 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      541 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      601 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
      661 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
      721 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
      781 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
      841 gatagtcttg taaatgcggg ccaagatcga tctgcacact ggtatttcgg tttttggggc
      901 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      961 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     1021 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1081 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1141 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1201 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1261 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1321 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1381 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1441 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaattggcg aagctcagtt
     1501 aacggccatt gtggccatca agcttggtac cagatctgga tccgccgcca ccatgagcct
     1561 gaacagcagc ttgagttgtc gcaaagagct tagtaatctg actgaagaag agggaggtga
     1621 agggggagta atcattacac agttcatagc tattatagtc ataaccattt tcgtttgcct
     1681 cgggaatctc gtcatagtag ttacactgta caaaaaatct tacttgctta ccctgtccaa
     1741 caagtttgta ttttccctga ccctctctaa cttccttctt tctgtgcttg tactcccctt
     1801 cgtagtaacg tcttcaataa ggagggagtg gattttcgga gtagtatggt gcaatttttc
     1861 agcgctgctg tatctgctta taagtagcgc aagtatgctt acgctcgggg ttatcgccat
     1921 tgatcgatat tacgcggtat tgtacccaat ggtataccct atgaaaataa cgggaaaccg
     1981 agccgtcatg gcactcgttt acatttggct ccactcactc attggctgtc tcccgccgct
     2041 gttcgggtgg agcagtgtcg agttcgacga atttaagtgg atgtgcgtcg cggcttggca
     2101 ccgggagcca ggctatacgg ccttttggca aatttggtgc gctttgttcc cgtttttggt
     2161 aatgctggtg tgttatggct ttatcttccg cgtggcgcga gtcaaagccc gaaaggttca
     2221 ttgcggaacg gttgtcattg tagaggaaga tgctcaacgc acgggccgga aaaactcttc
     2281 taccagcacg tcatcttctg gcagtcgacg gaatgcattc caaggggtag tgtactccgc
     2341 caaccaatgc aaggctctga tcacgattct ggtggtactt ggcgcgttca tggtgacctg
     2401 ggggccgtat atggtcgtta ttgcgtccga ggctctctgg ggaaagtctt cagtatctcc
     2461 cagcctggaa acttgggcaa cgtggctttc tttcgcgtcc gctgtctgcc atccattgat
     2521 ttatggattg tggaataaaa ccgtccgaaa ggaattgctt ggtatgtgct tcggtgatcg
     2581 ctattacaga gaaccctttg tgcagcgaca gcggaccagc cgattgtttt ctataagcaa
     2641 tcggattaca gaccttgggc tgtccccaca ccttaccgcg cttatggctg gagggcaacc
     2701 cttggggcat tcaagttcta ctggtgacac gggcttttca tgttcccagg attcaggtac
     2761 tgatatgatg cttctggaag actacacgtc cgacgataat ccccccagtc attgtacatg
     2821 tccgcccaaa cgccgatcct ccgtgacttt tgaggatgaa gtggagcaaa taaaagaagc
     2881 tgctaagaac agcattctgc atgttaaagc tgaggtccac aagtcactcg atagctatgc
     2941 ggctagcctc gccaaggcga tcgaagcaga ggccaagatc aacctgttcg gcgaagaggc
     3001 gctcccaggc gtcctggtaa cagctcggac cgttccaggt gggggcttcg gtggtagacg
     3061 cggcagcaga actctggtct cccagaggct gcagctgcaa agcattgaag aaggcgatgt
     3121 gctcgcagcc gaacaacggg aattctaccc atacgatgtt cctgactatg cgtaggcggc
     3181 cgcactcgag atatcggcct ccagggcctc tagagggccc tattctatag tgtcacctaa
     3241 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3301 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3361 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3421 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3481 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3541 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3601 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3661 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3721 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3781 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3841 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3901 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3961 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4141 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4201 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4261 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4321 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4381 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4441 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4501 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4561 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4621 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4681 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4741 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4801 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4861 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4921 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4981 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5041 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5101 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5161 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5221 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5281 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5341 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5401 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5461 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5581 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5641 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5701 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5761 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5821 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
