LOCUS       sequence-497389-        7242 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  Expression of Galphas (GNAS) wild type EE-tagged (internal).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7242)
  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 7242)
  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 GNAS EE WT
FEATURES             Location/Qualifiers
     source          1..7242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(253..357)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(436..455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        686..1868
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          917..1859
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1816..1836
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1941..1950
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3183..3201)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3184..3201)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3221..3238)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3227..3451
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3497..3925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3584..3603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3794..3815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3934..3954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3939..4268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4119..4254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4181..4200
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4335..5129
                     /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(4389..4408)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4999..5018
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5303..5424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5340..5359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5394..5413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5473..5489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5473..5489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5486..5508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5497..5513
                     /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(5521..5527,5528..5545,5546..5551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5566..5587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5704..5721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5875..6463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5955..5974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6634..7242,1..183,184..252))
                     /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 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
       61 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
      241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
      301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      361 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgatctc ccgatcccct
      421 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
      481 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
      541 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
      601 agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt
      661 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc
      721 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      781 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      841 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      901 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
      961 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
     1021 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
     1081 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
     1141 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
     1201 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatcgatc
     1261 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
     1321 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
     1381 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
     1441 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
     1501 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
     1561 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
     1621 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     1681 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1741 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1801 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1861 tgtcgtgagg aattggcgaa gctcagttaa cggccattgt ggccatcaag cttggtacca
     1921 gatctggatc cgctagcgcc gccaccatgg gctgcctcgg gaacagtaag accgaggacc
     1981 agcgcaacga ggagaaggcg cagcgtgagg ccaacaaaaa gatcgagaag cagctgcaga
     2041 aggacaagca ggtctaccgg gccacgcacc gcctgctcct tttgggcgct ggagaatctg
     2101 gtaaaagcac cattgtgaag cagatgagga tcctgcatgt taatgggttt aatggagagg
     2161 gcggcgaaga ggacccgcag gctgcaagga gcaacagcga tggtgagaag gcaaccaaag
     2221 tgcaggacat caaaaacaac ctgaaagagg cgattgaaac cattgtggcc gccatgagca
     2281 acctggtgcc ccccgtggag ctggccaacc ccgagaacca gttcagagtg gactacatcc
     2341 tgagtgtgat gaacgtgcct gactttgact tccctcccga attctatgag catgccaagg
     2401 ctctgtggga ggatgaagga gtgcgtgcct gctacgaacg ctccaacgag taccagctga
     2461 ttgactgtgc ccagtacttc ctggacaaga tcgacgtgat caagcaggct gaatatatgc
     2521 cgaccgagca ggacctgctt cgctgccgtg tcctgacttc tggaatcttt gagaccaagt
     2581 tccaggtgga caaagtcaac ttccacatgt ttgacgtggg tggccagcgc gatgaacgcc
     2641 gcaagtggat ccagtgcttc aacgatgtga ctgccatcat cttcgtggtg gccagcagca
     2701 gctacaacat ggtcatccgg gaggacaacc agaccaaccg cctgcaggag gctctgaacc
     2761 tcttcaagag catctggaac aacagatggc tgcgcaccat ctctgtgatc ctgttcctca
     2821 acaagcaaga tctgctcgct gagaaagtcc ttgctgggaa atcgaagatt gaggactact
     2881 ttccagaatt tgctcgctac actactcctg aggatgctac tcccgagccc ggagaggacc
     2941 cacgcgtgac ccgggccaag tacttcattc gagatgagtt tctgaggatc agcactgcca
     3001 gtggagatgg gcgtcactac tgctaccctc atttcacctg cgctgtggac actgagaaca
     3061 tccgccgtgt gttcaacgac tgccgtgaca tcattcagcg catgcacctt cgtcagtacg
     3121 agctgctcta ataggcggcc gcactcgaga tatcggcctc cagggcctct agagggccct
     3181 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
     3241 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3301 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3361 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3421 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     3481 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3541 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3601 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3661 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3721 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3781 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3841 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3901 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3961 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4021 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4081 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4141 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4201 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4261 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4321 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4381 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4441 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4501 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4561 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4621 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4681 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4741 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4801 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4861 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4921 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4981 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5041 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5101 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5161 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5221 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5281 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5341 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5401 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5521 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc cc
//
