LOCUS       sequence-173605-        7202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human DAGLb protein with a HA tag inserted in the first 
            extracellular loop, and intracellular V5 tag located on the 
            C-terminal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7202)
  TITLE     Unpublished Doherty Lab Plasmids
REFERENCE   2  (bases 1 to 7202)
  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"
COMMENT     Sequence Label: HA-hDAGLb-V5
FEATURES             Location/Qualifiers
     source          1..7202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(3009..3033)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3059..3100
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3140..3159)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3184..3232
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3434..3862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3521..3540)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3731..3752
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3871..3891)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3876..4205
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4056..4191
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4118..4137
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4253..4300
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4319..4717
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4875..4996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4912..4931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4966..4985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5045..5061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5045..5061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5058..5080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5069..5085
                     /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(5093..5099,5100..5117,5118..5123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5276..5293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5447..6035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5527..5546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6206..6997,6998..7066))
                     /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     6829..6848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7067..7171)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggcttcacc atgccgggga tggtactctt
      961 cggccggcgc tgggccatcg ccagcgacga cttggtcttc ccagggttct tcgagctggt
     1021 cgtgcgagtg ctgtggtgga ttggcattct gacgttgtat ctcatgcaca gaggaaagct
     1081 ggacggaagc ggaagctatc catatgacgt cccagactat gccggaagcg gaagctgtgc
     1141 tggtggagcc ttgctcagca gttacttgat cgtcctcatg attctcctgg cagttgtcat
     1201 atgtactgtg tcagccatca tgtgtgtcag catgagagga acgatttgta accctggacc
     1261 gcggaagtct atgtctaagc tgctttacat ccgcctggcg ctgttttttc cagagatggt
     1321 ctgggcctct ctgggggctg cctgggtggc agatggtgtt cagtgcgaca ggacagttgt
     1381 aaacggcatc atcgcaaccg tcgtggtcag ttggatcatc atcgctgcca cagtggtttc
     1441 cattatcatt gtctttgacc ctcttggggg gaaaatggct ccatattcct ctgccggccc
     1501 cagccacctg gatagtcatg attcaagcca gttacttaat ggcctcaaga cagcagctac
     1561 aagcgtgtgg gaaaccagaa tcaagctctt gtgctgttgc attgggaaag acgaccatac
     1621 tcgggttgct ttttcgagta cggcagagct tttctcaacc tacttttcag acacagatct
     1681 ggtgcccagc gacattgcgg cgggcctcgc cctgcttcat cagcaacagg acaatatcag
     1741 gaacaaccaa gagcctgccc aggtggtctg ccatgcccca gggagctccc aggaagctga
     1801 tctggatgca gaattagaaa actgccatca ttacatgcag tttgcagcag cggcctatgg
     1861 gtggcccctc tacatctaca gaaaccccct cacggggctg tgcaggattg gtggtgactg
     1921 ctgcagaagc agaaccacag actatgactt ggtcggaggc gatcagctca actgtcactt
     1981 cggctccatc ctgcacacca cagggctgca gtacagggac ttcatccacg tcagcttcca
     2041 tgacaaggtt tacgagctgc cgtttttagt ggctctggat cacaggaaag agtctgttgt
     2101 ggtcgctgtg agggggacca tgtctctgca ggatgtcctt acggacctgt cagcggagag
     2161 tgaggtgctg gacgtggagt gtgaggtgca ggaccgcctg gcacacaagg gtatttctca
     2221 agctgccaga tacgtttacc aacgactcat caacgacggg attttgagcc aagccttcag
     2281 cattgctcct gagtaccggc tggtcatagt gggccacagc ctcgggggag gggcggccgc
     2341 cctgctggcc accatgctca gagccgccta cccgcaggtc aggtgctacg ccttctcccc
     2401 accccggggg ctgtggagca aagctctgca ggaatattct cagagcttca tcgtgtcact
     2461 cgtcctgggg aaggatgtga ttcccaggct cagtgtgacc aacttggaag atctgaagag
     2521 aagaatcttg cgagtggtcg cgcactgcaa taaacccaag tacaagatct tgctgcacgg
     2581 tttgtggtac gaactgtttg gaggaaaccc caacaacttg cccacggagc tggacggggg
     2641 cgaccaggaa gtcctgacac agcctcttct gggggagcag agcctactga cgcgctggtc
     2701 cccggcctac agcttctcca gcgactcccc actggactct tctcccaagt acccccctct
     2761 ctaccctccc ggcaggatca tccacctgca ggaggagggc gcctcggggc ggtttggctg
     2821 ctgctctgct gctcactata gcgccaagtg gtcacacgaa gcggaattca gcaaaatact
     2881 cataggtccg aagatgctca ccgaccacat gccagacatc ctgatgcggg ccttggacag
     2941 cgtggtctcc gacagagcgg cctgcgtctc ctgtccagca cgaggggtct ccagtgtgga
     3001 cgtggcccac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg
     3061 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt
     3121 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3181 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3241 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3301 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3361 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3421 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3481 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3541 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3601 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3661 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3721 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3781 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3841 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4081 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4141 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4201 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
     4261 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
     4321 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     4381 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
     4441 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     4501 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     4561 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     4621 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     4681 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
     4741 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4801 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4861 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4921 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4981 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7201 tc
//
