LOCUS       sequence-501599-        6900 bp    DNA     circular SYN 03-MAR-2026
DEFINITION  Expressing Venus funsed human G_s Q227L, short isoform.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  TITLE     Garcia-Marcos Lab Plasmids 2025
REFERENCE   2  (bases 1 to 6900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 3, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pcDNA3.1-G-alpha-s 99-Venus Q227L (short)
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(98..202)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(277..296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        468..847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        848..1051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1002..1022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1096..1115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1096..1114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1537..1539,1540..2250)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1579..1600)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1840..1859)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2187..2208
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3141..3158)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3147..3371
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3417..3845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3504..3523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3714..3735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3854..3874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3859..4188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4039..4174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4101..4120
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4236..4283
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4302..4676
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4806..4927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4843..4862)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4897..4916
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4976..4992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4976..4992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4989..5011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5000..5016
                     /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(5024..5030,5031..5048,5049..5054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5069..5090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5207..5224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5378..5966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5458..5477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6137..6900,1..28,29..97))
                     /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     6760..6779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
       61 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
      121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
      181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
      241 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
      301 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
      361 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
      421 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
      481 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      541 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      601 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      661 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      721 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      781 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      841 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      901 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      961 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     1021 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     1081 ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct agcgtttaaa
     1141 cgggccctct agactcgagc ggccgccact gtgctggata tctgcagaat tccaccacac
     1201 tggactagtg gatccgagct cggtaccatg ggctgcctcg ggaacagtaa gaccgaggac
     1261 cagcgcaacg aggagaaggc gcagcgtgag gccaacaaaa agatcgagaa gcagctgcag
     1321 aaggacaagc aggtctaccg ggccacgcac cgcctgctgc tgctgggtgc tggagaatct
     1381 ggtaaaagca ccattgtgaa gcagatgagg atcctgcatg ttaatgggtt taatggagac
     1441 agtgagaagg caaccaaagt gcaggacatc aaaaacaacc tgaaagaggc gattgaaacc
     1501 attgtggccg ccatgagcaa cctgggtggc ggaggggtga gcaagggcga ggagctgttc
     1561 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1621 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     1681 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
     1741 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1801 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1861 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1921 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1981 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2041 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     2101 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
     2161 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2221 atcactctcg gcatggacga gctgtacaag ggcggtgggg gagtgccccc cgtggagctg
     2281 gccaaccccg agaaccagtt cagagtggac tacattctga gtgtgatgaa cgtgcctgac
     2341 tttgacttcc ctcccgaatt ctatgagcat gccaaggctc tgtgggagga tgaaggagtg
     2401 cgtgcctgct acgaacgctc caacgagtac cagctgattg actgtgccca gtacttcctg
     2461 gacaagatcg acgtgatcaa gcaggctgac tatgtgccga gcgatcagga cctgcttcgc
     2521 tgccgtgtcc tgacttctgg aatctttgag accaagttcc aggtggacaa agtcaacttc
     2581 cacatgtttg acgtgggtgg cctgcgcgat gaacgccgca agtggatcca gtgcttcaac
     2641 gatgtgactg ccatcatctt cgtggtggcc agcagcagct acaacatggt catccgggag
     2701 gacaaccaga ccaaccgcct gcaggaggct ctgaacctct tcaagagcat ctggaacaac
     2761 agatggctgc gcaccatctc tgtgatcctg ttcctcaaca agcaagatct gctcgctgag
     2821 aaagtccttg ctgggaaatc gaagattgag gactactttc cagaatttgc tcgctacact
     2881 actcctgagg atgctactcc cgagcccgga gaggacccac gcgtgacccg ggccaagtac
     2941 ttcattcgag atgagtttct gaggatcagc actgccagtg gagatgggcg tcactactgc
     3001 taccctcatt tcacctgcgc tgtggacact gagaacatcc gccgtgtgtt caacgactgc
     3061 cgtgacatca ttcagcgcat gcaccttcgt cagtacgagc tgctataagg taccaagctt
     3121 aagtttaaac cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3181 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3241 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3301 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3361 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3421 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3481 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3541 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3601 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3661 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3721 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3781 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3841 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4201 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     4261 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     4321 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     4381 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     4441 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     4501 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     4561 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     4621 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     4681 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4741 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4801 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4861 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4921 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4981 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5041 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5101 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5161 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5221 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5281 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5341 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5401 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5461 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5521 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5581 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5641 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5701 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5761 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5821 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5881 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5941 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6001 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6061 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6121 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6361 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6421 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6661 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6781 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6841 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
//
