LOCUS       62056_18350        8263 bp DNA     circular SYN 03-JAN-2021
DEFINITION  Targeting vector pPB-B2-TARG-GE, complete sequence.
ACCESSION   MT917127
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8263)
  AUTHORS   Kondrashov A, Mohd Yusof NAN., Hasan A, Goulding J, Kodagoda T, 
            Hoang DM, Vo NTN., Melarangi T, Dolatshad N, Gorelik J, Hill SJ, 
            Harding SE, Denning C.
  TITLE     CRISPR/Cas9-mediated generation and analysis of N terminus 
            polymorphic models of beta2AR in isogenic hPSC-derived 
            cardiomyocytes
  JOURNAL   Mol Ther Methods Clin Dev 20, 39-53 (2020)
  PUBMED    33335946
REFERENCE   2  (bases 1 to 8263)
  AUTHORS   Kondrashov A, Mohd Yusof NAN., Hasan A, Goulding J, Kodagoda T, Minh
            DH, Thi NV, Melarangi T, Dolatshad N, Gorelik J, Hill S, Harding SE,
            Denning C.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 8263)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Methods Clin Dev 20, 39-53 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-AUG-2020) School of Medicine, Division of Cancer "
FEATURES             Location/Qualifiers
     source          1..8263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            698..1731
                     /gene="adrB2"
                     /label=adrB2
                     /note="5' homology arm"
     mobile_element  1732..2059
                     /mobile_element_type="transposon"
                     /label=transposon
                     /note="PiggyBac 5' terminal repeat domain"
     protein_bind    2060..2084
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2179..2678
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        2690..2737
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2762..3355
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     CDS             3362..4354
                     /label=Delta-TK
                     /note="truncated herpes simplex virus thymidine kinase
                     (Salomon et al., 1995)"
     polyA_signal    4385..4609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4695..4719)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     mobile_element  4720..4967
                     /mobile_element_type="transposon"
                     /label=transposon
                     /note="PiggyBac 3' terminal repeat domain"
     gene            4968..5999
                     /gene="adrB2"
                     /label=adrB2
                     /note="3' homology arm"
     primer_bind     complement(6022..6038)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6075..6093)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6114..6130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6138..6154)
                     /label=lac operator
                     /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(6162..6192)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6207..6228)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6516..7104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7278..8135)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8136..8240)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatttc ggagtaccca gatggagaca
      721 tccgtgtctg tgtcgctctg gatgcctcca agccagcgtg tgtttacttt ctgtgtgtgt
      781 caccatgtct ttgtgcttct gggtgcttct gtgtttgttt ctggccgcgt ttctgtgttg
      841 gacaggggtg actttgtgcc ggatggcttc tgtgtgagag cgcgcgcgag tgtgcatgtc
      901 ggtgagctgg gagggtgtgt ctcagtgtct atggctgtgg ttcggtataa gtctgagcat
      961 gtctgccagg gtgtatttgt gcctgtatgt gcgtgcctcg gtgggcactc tcgtttcctt
     1021 ccgaatgtgg ggcagtgccg gtgtgctgcc ctctgccttg agacctcaag ccgcgcaggc
     1081 gcccagggca ggcaggtagc ggccacagaa gagccaaaag ctcccgggtt ggctggtaag
     1141 gacaccacct ccagctttag ccctctgggg ccagccaggg tagccgggaa gcagtggtgg
     1201 cccgccctcc agggagcagt tgggccccgc ccgggccagc cccaggagaa ggagggcgag
     1261 gggaggggag ggaaagggga ggagtgcctc gccccttcgc ggctgccggc gtgccattgg
     1321 ccgaaagttc ccgtacgtca cggcgagggc agttccccta aagtcctgtg cacataacgg
     1381 gcagaacgca ctgcgaagcg gcttcttcag agcacgggct ggaactggca ggcaccgcga
     1441 gcccctagca cccgacaagc tgagtgtgca ggacgagtcc ccaccacacc cacaccacag
     1501 ccgctgaatg aggcttccag gcgtccgctc gcggcccgca gagccccgcc gtgggtccgc
     1561 ccgctgaggc gcccccagcc agtgcgctca cctgccagac tgcgcgccat ggggcaaccc
     1621 gggaacggca gcgccttctt gctggcaccc aatggaagcc atgcgccgga ccacgacgtc
     1681 acgcaggaaa gggacgaggt gtgggtggtg ggcatgggca tcgtcatgtc tttaacccta
     1741 gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa atattgctct ctctttctaa
     1801 atagcgcgaa tccgtcgctg tgcatttagg acatctcagt cgccgcttgg agctcccgtg
     1861 aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa ctataacgac cgcgtgagtc
     1921 aaaatgacgc atgattatct tttacgtgac ttttaagatt taactcatac gataattata
     1981 ttgttatttc atgttctact tacgtgataa cttattatat atatattttc ttgttataga
     2041 tatcaactag aatgctagca caagtttgta caaaaaagca ggctggcgcc ggaaccaatt
     2101 cagtcgactg gatccggtac cgggcccccc ctcgaggtcg agacggtatc gataagcttg
     2161 atatcgaata attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     2221 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     2281 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     2341 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     2401 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     2461 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     2521 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     2581 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     2641 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagcctg ttgacaatta
     2701 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggg
     2761 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     2821 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     2881 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     2941 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3001 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3061 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3121 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     3181 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     3241 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     3301 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccggatc
     3361 catgcccacg ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac
     3421 cacgcaactg ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat
     3481 gacttactgg caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca
     3541 ccgcctcgac cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca
     3601 gataacaatg ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg
     3661 ggaggctggg agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc
     3721 catcgccgcc ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca
     3781 ggccgtgctg gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt
     3841 gggggccctt ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga
     3901 gcggcttgac ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac
     3961 ggtgcggtat ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac
     4021 ggccgtgccg ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg
     4081 ggacacgtta tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta
     4141 caacgtgttt gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt
     4201 tatcctggat tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc
     4261 cgggatggtc cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc
     4321 gcgcacgttt gcccgggaga tgggggaggc taactgagct ctagagctcg ctgatcagcc
     4381 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4441 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4501 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4561 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4621 gaaagaacca gctggggctc gagatccact agttctagcc tcgaggctag agcggccgca
     4681 ctcgagatat ctagacccag ctttcttgta caaagtggta ctagttaaaa gttttgttac
     4741 tttatagaag aaattttgag tttttgtttt tttttaataa ataaataaac ataaataaat
     4801 tgtttgttga atttattatt agtatgtaag tgtaaatata ataaaactta atatctattc
     4861 aaattaataa ataaacctcg atatacagac cgataaaaca catgcgtcaa ttttacgcat
     4921 gattatcttt aacgtacgtc acaatatgat tatctttcta gggttaatcg tcctggccat
     4981 cgtgtttggc aatgtgctgg tcatcacagc cattgccaag ttcgagcgtc tgcagacggt
     5041 caccaactac ttcatcactt cactggcctg tgctgatctg gtcatgggcc tggcagtggt
     5101 gccctttggg gccgcccata ttcttatgaa aatgtggact tttggcaact tctggtgcga
     5161 gttttggact tccattgatg tgctgtgcgt cacggccagc attgagaccc tgtgcgtgat
     5221 cgcagtggat cgctactttg ccattacttc acctttcaag taccagagcc tgctgaccaa
     5281 gaataaggcc cgggtgatca ttctgatggt gtggattgtg tcaggcctta cctccttctt
     5341 gcccattcag atgcactggt accgggccac ccaccaggaa gccatcaact gctatgccaa
     5401 tgagacctgc tgtgacttct tcacgaacca agcctatgcc attgcctctt ccatcgtgtc
     5461 cttctacgtt cccctggtga tcatggtctt cgtctactcc agggtctttc aggaggccaa
     5521 aaggcagctc cagaagattg acaaatctga gggccgcttc catgtccaga accttagcca
     5581 ggtggagcag gatgggcgga cggggcatgg actccgcaga tcttccaagt tctgcttgaa
     5641 ggagcacaaa gccctcaaga cgttaggcat catcatgggc actttcaccc tctgctggct
     5701 gcccttcttc atcgttaaca ttgtgcatgt gatccaggat aacctcatcc gtaaggaagt
     5761 ttacatcctc ctaaattgga taggctatgt caattctggt ttcaatcccc ttatctactg
     5821 ccggagccca gatttcagga ttgccttcca ggagcttctg tgcctgcgca ggtcttcttt
     5881 gaaggcctat gggaatggct actccagcaa cggcaacaca ggggagcaga gtggatatca
     5941 cgtggaacag gagaaagaaa ataaactgct gtgtgaagac ctcccaggca cggaagacta
     6001 tcgaattcct gcagcccggg ggatccacta gttctagagc ggccgccacc gcggtggagc
     6061 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     6121 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6181 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     6241 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     6301 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6361 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6421 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6481 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6541 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6601 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6661 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6721 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6781 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6841 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6901 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6961 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7021 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7081 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7141 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7201 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7261 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7321 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7381 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7441 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7501 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7561 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7621 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7681 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7741 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7801 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7861 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7921 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7981 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8041 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8101 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8161 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8221 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
