LOCUS       sequence-122192-        7299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G protein alpha-s internally tagged with HaloTag and EE epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  TITLE     Berlot Lab HaloTag Fusions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7299)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: G protein alpha-s-HaloTag
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(603..1061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(740..761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     955..974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1186..1204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1272..1376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1377..1445,1446..2237)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1595..1614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2310..2689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2690..2893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2844..2864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2938..2957
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2938..2956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3160..3169
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5571..5589)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5572..5589)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          5673..5738
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5868..5888
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6160..6294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6197..6216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6251..6270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7140..7275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
       61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
      601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
      841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
     1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
     1141 aagggatttt agacaggaac ggtacgccag ctggattacc aaagggcctc gtgatacgcc
     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2281 aacttcattt ttaatttctc tagcgcgttg acattgatta ttgactagtt attaatagta
     2341 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2401 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2461 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
     2521 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2581 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2641 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2701 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2761 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2821 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2881 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     2941 tacgactcac tatagggaga cccaagcttg gtcgaccaat tccgcagcag cagctcccgc
     3001 ggctcctgct ctgctccgcc tcggccccgg agcgaggggc ggagagccgc gcgcttgcct
     3061 tagtccgagc cgccaccctc cccgcgttcc ccgctttccc ggcccgcccg aggccgcccg
     3121 ccccgtcccc gccgccccgc agcccggccg cgccccgccg ccgccatggg ctgcctcggc
     3181 aacagtaaga ccgaggacca gcgcaacgag gagaaggcgc agcgcgaggc caacaaaaag
     3241 atcgagaagc agctgcagaa ggacaagcag gtctaccggg ccacgcaccg cctgctgctg
     3301 ctgggtgctg gagagtctgg caaaagcacc attgtgaagc agatgaggat tctacatgtt
     3361 aatgggttta acggagaggg atccggggga ggcgggagcg gtgggggagg cagtgaaatc
     3421 ggtacaggct tccccttcga cccccattat gtggaagtcc tgggcgagcg tatgcactac
     3481 gtcgatgttg gaccgcggga tggcacgcct gtgctgttcc tgcacggtaa cccgacctcg
     3541 tcctacctgt ggcgcaacat catcccgcat gtagcaccga gtcatcggtg cattgctcca
     3601 gacctgatcg ggatgggaaa atcggacaaa ccagacctcg attatttctt cgacgaccac
     3661 gtccgctacc tcgatgcctt catcgaagcc ttgggtttgg aagaggtcgt cctggtcatc
     3721 cacgactggg gctcagctct cggattccac tgggccaagc gcaatccgga acgggtcaaa
     3781 ggtattgcat gtatggaatt catccggcct atcccgacgt gggacgaatg gccagaattc
     3841 gcccgtgaga ccttccaggc cttccggacc gccgacgtcg gccgagagtt gatcatcgat
     3901 cagaacgctt tcatcgaggg tgcgctcccg atgggggtcg tccgtccgct tacggaggtc
     3961 gagatggacc actatcgcga gcccttcctc aagcctgttg accgagagcc actgtggcga
     4021 ttccccaacg agctgcccat cgccggtgag cccgcgaaca tcgtcgcgct cgtcgaggca
     4081 tacatgaact ggctgcacca gtcacctgtc ccgaagttgt tgttctgggg cacacccggc
     4141 gtactgatcc ccccggccga agccgcgaga cttgccgaaa gcctccccaa ctgcaagaca
     4201 gtggacatcg gcccgggatt gttcttgctc caggaagaca acccggacct tatcggcagt
     4261 gagatcgcgc gctggctccc cgggctggcc ggctcagggg gcggtggcag cgggggtggc
     4321 ggatccgatg gtgagaaggc caccaaagtg caggacatca aaaacaacct gaaggaggcc
     4381 attgaaacca ttgtggccgc catgagcaac ctggtgcccc ccgtggagct ggccaaccct
     4441 gagaaccagt tcagagtgga ctacattctg agcgtgatga acgtgccaaa ctttgacttc
     4501 ccacctgaat tctatgagca tgccaaggct ctgtgggagg atgagggagt tcgtgcctgc
     4561 tacgagcgct ccaacgagta ccagctgatc gactgtgccc agtacttcct ggacaagatt
     4621 gatgtgatca agcaggccga gtacatgccg actgagcagg acctgcttcg ctgccgcgtc
     4681 ctgacctctg gaatctttga gaccaagttc caggtggaca aagtcaactt ccacatgttc
     4741 gatgtgggcg gccagcgcga tgaacgccgc aagtggattc agtgcttcaa tgacgtaacg
     4801 gcaattatat tcgtggtggc cagcagcagc tacaacatgg tcatccggga ggacaaccag
     4861 accaaccgtc tgcaggaggc tctgaacctc ttcaagagca tctggaacaa cagatggctg
     4921 cgtaccatct ctgtgatcct cttcctcaac aagcaagatc tgcttgctga gaaggtcctc
     4981 gctgggaaat cgaagattga ggactacttt ccagagttcg ctcgctacac cactcctgag
     5041 gatgcgactc ccgagcccgg agaggaccca cgcgtgaccc gggccaagta cttcatccgg
     5101 gatgagtttc tgagaatcag cactgctagt ggagatggac gtcactactg ctaccctcac
     5161 tttacctgcg ccgtggacac tgagaacatc cgccgtgtct tcaacgactg ccgtgacatc
     5221 atccagcgca tgcatcttcg ccaatacgag ctgctctaag aagggaacgc ccaaatttaa
     5281 ttcagcctta agcacaatta attaagagtg aaacgcaatc gtacaagcag ttgatcaccc
     5341 accatagggc atgatcaaca ccgcaacctt tcccttttct ccccagtgat tctgaaaacc
     5401 ccctcttccc ttcagcttgc ttagatgttc caaatttagt aagctccaag cttggccaag
     5461 cttggtaccg agctcggatc gatccactag taacggccgc cagtgtgctg gaattctgca
     5521 gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     5581 cacctaaatg ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
     5641 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
     5701 gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg
     5761 aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc
     5821 catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga
     5881 gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg
     5941 tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac
     6001 tgctatacaa gaaaattatg gaaaaatatt tgatgtatag tgccttgact agagatcata
     6061 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     6121 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     6181 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6241 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcatcccg
     6301 ccatggtatc aacgccatat ttctatttac agtagggacc tcttcgttgt gtaggtaccg
     6361 ctgtattcct agggaaatag tagaggcacc ttgaactgtc tgcatcagcc atatagcccc
     6421 cgctgttcga cttacaaaca caggcacagt actgacaaac ccatacacct cctctgaaat
     6481 acccatagtt gctagggctg tctccgaact cattacaccc tccaaagtca gagctgtaat
     6541 ttcgccatca agggcagcga gggcttctcc agataaaata gcttctgccg agagtcccgt
     6601 aagggtagac acttcagcta atccctcgat gaggtctact agaatagtca gtgcggctcc
     6661 cattttgaaa attcacttac ttgatcagct tcagaagatg gcggagggcc tccaacacag
     6721 taattttcct cccgactctt aaaatagaaa atgtcaagtc agttaagcag gaagtggact
     6781 aactgacgca gctggccgtg cgacatcctc ttttaattag ttgctaggca acgccctcca
     6841 gagggcgtgt ggttttgcaa gaggaagcaa aagcctctcc acccaggcct agaatgtttc
     6901 cacccaatca ttactatgac aacagctgtt ttttttagta ttaagcagag gccggggacc
     6961 cctgggcccg cttactctgg agaaaaagaa gagaggcatt gtagaggctt ccagaggcaa
     7021 cttgtcaaaa caggactgct tctatttctg tcacactgtc tggccctgtc acaaggtcca
     7081 gcacctccat acccccttta ataagcagtt tgggaacggg tgcgggtctt actccgccca
     7141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     7201 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     7261 gcttttttgg aggcctaggc ttttgcaaaa agctaattc
//
