LOCUS       sequence-497391-        7137 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  Expression of PKA (PRKACA) wild type.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  AUTHORS   Krantz S, Bell BA, Lund K, Salinas Parra N, Ng Y, De Oliveira Rosa 
            N, Mukhopadhyay S, St Croix B, Sarin KY, Weigert R, Raimondi F, 
            Iglesias-Bartolome R
  TITLE     Dissection of Galphas and Hedgehog Signaling Crosstalk Reveals 
            Therapeutic Opportunities to Target Hedgehog-Dependent Tumors.
  JOURNAL   Cancer Res. 2025 Dec 29. doi: 10.1158/0008-5472.CAN-25-1109.
  PUBMED    41460725
REFERENCE   2  (bases 1 to 7137)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cancer Res.
            2025 Dec 29. doi: 10.1158/0008-5472.CAN-25-1109."
COMMENT     Sequence Label: pCEFL PRKACA HA WT
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(252..356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(435..454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        685..1867
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          916..1858
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1815..1835
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1934..1943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2999..3025
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3077..3095)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3078..3095)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3115..3132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3121..3345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3391..3819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3478..3497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3688..3709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3828..3848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3833..4162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4075..4094
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4229..5023
                     /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(4283..4302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4893..4912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5197..5318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5234..5253)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5288..5307
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5367..5383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5367..5383)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5380..5402)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5391..5407
                     /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(5415..5421,5422..5439,5440..5445))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5460..5481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5598..5615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5769..6357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5849..5868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6528..7137,1..182,183..251))
                     /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"
ORIGIN
        1 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
       61 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      121 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      181 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      241 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      301 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      361 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
      421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
      481 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
      541 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcga
      601 gctttgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc
      661 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
      721 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
      781 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
      841 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
      901 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
      961 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     1021 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     1081 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     1141 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     1201 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatcgatct
     1261 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
     1321 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     1381 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     1441 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     1501 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     1561 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     1621 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     1681 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     1741 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     1801 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     1861 gtcgtgagga attggcgaag ctcagttaac ggccattgtg gccatcaagc ttggtaccag
     1921 atctggatcc gccgccacca tgggcaacgc cgccgccgcc aagaagggca gcgagcagga
     1981 gagcgtgaaa gaattcttag ccaaagccaa agaagatttt cttaaaaaat gggaaagtcc
     2041 cgctcagaac acagcccact tggatcagtt tgaacgaatc aagaccctcg gcacgggctc
     2101 cttcgggcgg gtgatgctgg tgaaacacaa ggagaccggg aaccactatg ccatgaagat
     2161 cctcgacaaa cagaaggtgg tgaaactgaa acagatcgaa cacaccctga atgaaaagcg
     2221 catcctgcaa gctgtcaact ttccgttcct cgtcaaactc gagttctcct tcaaggacaa
     2281 ctcaaactta tacatggtca tggagtacgt gcccggcggg gagatgttct cacacctacg
     2341 gcggatcgga aggttcagtg agccccatgc ccgtttctac gcggcccaga tcgtcctgac
     2401 ctttgagtat ctgcactcgc tggatctcat ctacagggac ctgaagccgg agaatctgct
     2461 cattgaccag cagggctaca ttcaggtgac agacttcggt ttcgccaagc gcgtgaaggg
     2521 ccgcacttgg accttgtgcg gcacccctga gtacctggcc cctgagatta tcctgagcaa
     2581 aggctacaac aaggccgtgg actggtgggc cctgggggtt cttatctatg aaatggccgc
     2641 tggctacccg cccttcttcg cagaccagcc catccagatc tatgagaaga tcgtctctgg
     2701 gaaggtgcgc ttcccttccc acttcagctc tgacttgaag gacctgctgc ggaacctcct
     2761 gcaggtagat ctcaccaagc gctttgggaa cctcaagaat ggggtcaacg atatcaagaa
     2821 ccacaagtgg tttgccacaa ctgactggat tgccatctac cagaggaagg tggaagctcc
     2881 cttcatacca aagtttaaag gccctgggga tacgagtaac tttgacgact atgaggaaga
     2941 agaaatccgg gtctccatca atgagaagtg tggcaaggag ttttctgagt ttgctagcta
     3001 cccatacgac gtgccagatt acgcctaggc ggccgcactc gagatatcgg cctccagggc
     3061 ctctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     3121 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3181 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3241 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3301 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3361 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3781 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4201 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4261 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4321 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4381 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4441 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4501 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4561 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4621 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4681 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4741 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4801 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4861 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4921 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4981 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5041 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5101 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5161 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5221 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5281 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5341 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5401 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5461 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5521 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5581 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5641 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5701 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5761 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5821 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5881 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5941 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6001 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6061 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccg
//
