LOCUS       sequence-115422-        6286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A2A-adenosine receptor tagged with HaloTag on amino terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6286)
  TITLE     Berlot Lab HaloTag Fusions
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6286)
  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            "Unpublished"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: HaloTag-A2A-adenosine receptor
FEATURES             Location/Qualifiers
     source          1..6286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     718..742
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1072..1081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1078..1155
                     /codon_start=1
                     /product="human growth hormone signal sequence"
                     /label=hGH signal sequence
                     /translation="MATGSRTSLLLAFGLLCLPWLQEGSA"
     polyA_signal    3395..3516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3406..3425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3460..3479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3702..4157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3789..3808)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3999..4020
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4224..4243)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4343..4365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4403..4421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4594..4662,4663..5454)
                     /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     complement(4812..4831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5625..6213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6114..6133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagccag ctggcgcgga tgccaccatg
     1081 gctacaggct cccggacgtc cctgctcctg gcttttggcc tgctctgcct gccctggctt
     1141 caagagggca gtgccttccc aaccattccc ttatccggat ccgaaatcgg tacaggcttc
     1201 cccttcgacc cccattatgt ggaagtcctg ggcgagcgta tgcactacgt cgatgttgga
     1261 ccgcgggatg gcacgcctgt gctgttcctg cacggtaacc cgacctcgtc ctacctgtgg
     1321 cgcaacatca tcccgcatgt agcaccgagt catcggtgca ttgctccaga cctgatcggg
     1381 atgggaaaat cggacaaacc agacctcgat tatttcttcg acgaccacgt ccgctacctc
     1441 gatgccttca tcgaagcctt gggtttggaa gaggtcgtcc tggtcatcca cgactggggc
     1501 tcagctctcg gattccactg ggccaagcgc aatccggaac gggtcaaagg tattgcatgt
     1561 atggaattca tccggcctat cccgacgtgg gacgaatggc cagaattcgc ccgtgagacc
     1621 ttccaggcct tccggaccgc cgacgtcggc cgagagttga tcatcgatca gaacgctttc
     1681 atcgagggtg cgctcccgat gggggtcgtc cgtccgctta cggaggtcga gatggaccac
     1741 tatcgcgagc ccttcctcaa gcctgttgac cgagagccac tgtggcgatt ccccaacgag
     1801 ctgcccatcg ccggtgagcc cgcgaacatc gtcgcgctcg tcgaggcata catgaactgg
     1861 ctgcaccagt cacctgtccc gaagttgttg ttctggggca cacccggcgt actgatcccc
     1921 ccggccgaag ccgcgagact tgccgaaagc ctccccaact gcaagacagt ggacatcggc
     1981 ccgggattgt tcttgctcca ggaagacaac ccggacctta tcggcagtga gatcgcgcgc
     2041 tggctccccg ggctggccgg ctccggtggg ggcgggagca tgcccatcat gggctcctcg
     2101 gtgtacatca cggtggagct ggccattgct gtgctggcca tcctgggcaa tgtgctggtg
     2161 tgctgggccg tgtggctcaa cagcaacctg cagaacgtca ccaactactt tgtggtgtca
     2221 ctggcggcgg ccgacatcgc agtgggtgtg ctcgccatcc cctttgccat caccatcagc
     2281 accgggttct gcgctgcctg ccacggctgc ctcttcattg cctgcttcgt cctggtcctc
     2341 acgcagagct ccatcttcag tctcctggcc atcgccattg accgctacat tgccatccgc
     2401 atcccgctcc ggtacaatgg cttggtgacc ggcacgaggg ctaagggcat cattgccatc
     2461 tgctgggtgc tgtcgtttgc catcggcctg actcccatgc taggttggaa caactgcggt
     2521 cagccaaagg agggcaagaa ccactcccag ggctgcgggg agggccaagt ggcctgtctc
     2581 tttgaggatg tggtccccat gaactacatg gtgtacttca acttctttgc ctgtgtgctg
     2641 gtgcccctgc tgctcatgct gggtgtctat ttgcggatct tcctggcggc gcgacgacag
     2701 ctgaagcaga tggagagcca gcctctgccg ggggagcggg cacggtccac actgcagaag
     2761 gaggtccatg ctgccaagtc actggccatc attgtggggc tctttgccct ctgctggctg
     2821 cccctacaca tcatcaactg cttcactttc ttctgccccg actgcagcca cgcccctctc
     2881 tggctcatgt acctggccat cgtcctctcc cacaccaatt cggttgtgaa tcccttcatc
     2941 tacgcctacc gtatccgcga gttccgccag accttccgca agatcattcg cagccacgtc
     3001 ctgaggcagc aagaaccttt caaggcagct ggcaccagtg cccgggtctt ggcagctcat
     3061 ggcagtgacg gagagcaggt cagcctccgt ctcaacggcc acccgccagg agtgtgggcc
     3121 aacggcagtg ctccccaccc tgagcggagg cccaatggct acgccctggg gctggtgagt
     3181 ggagggagtg cccaagagtc ccaggggaac acgggcctcc cagacgtgga gctccttagc
     3241 catgagctca agggagtgtg cccagagccc cctggcctag atgaccccct ggcccaggat
     3301 ggagcaggag tgtcctgagg atccgccggc taatagttaa ttaagtaggc ggccgcttcc
     3361 ctttagtgag ggttaatgct tcgagcagac atgataagat acattgatga gtttggacaa
     3421 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     3481 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     3541 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     3601 tgtggtaaaa tccgataagg atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg
     3661 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     3721 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3781 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgcgg gctttccccg
     3841 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3901 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3961 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4021 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4081 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4141 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4201 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4261 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4321 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4381 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4441 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6241 ttcctggcct tttgctggcc ttttgctcac atggctcgac agatct
//
