LOCUS       sequence-426611-        7007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  barcoded assay,  internal control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7007)
  AUTHORS   Popovic L, Rossner MJ, Wehr MC
  TITLE     Protocol for identifying properties of ERBB receptor antagonists 
            using the barcoded ERBBprofiler assay.
  JOURNAL   STAR Protoc. 2024 Apr 16;5(2):102987. doi: 
            10.1016/j.xpro.2024.102987.
  PUBMED    38635397
REFERENCE   2  (bases 1 to 7007)
  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: "STAR 
            Protoc."; date: "2024-04-16"; volume: "5(2):102987. doi"; pages: " 
            10.1016/j.xpro.2024.102987"
COMMENT     Sequence Label: pAAV_MLP-BC0250-luc2
FEATURES             Location/Qualifiers
     source          1..7007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     primer_bind     complement(200..219)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    484..508
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(554..575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(709..729)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    870..890
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        917..935
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(998..1017)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(999..1017)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1041..1065)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1144..1153
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1150..2802
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTITEKEIVDYVASQATTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    2830..3418
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2883..2903)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3301..3312)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3445..3462)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3451..3675
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3885..4214)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3899..4034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4199..4219
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4460..5048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4540..4559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5219..6010,6011..6079))
                     /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     5842..5861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6080..6184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6252..6270
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6308..6330)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6430..6449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6663..6682
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6972..6990
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6972..6989
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      241 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      301 catcctgggg cacaagctgg agtacaacta caacagccac aacgtccata tcatggccga
      361 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      421 cgtgcagctc gccgaccact catatgtact cgagttgtcg acggtatcga taagcttgat
      481 atcacaagtt tgtacaaaaa agcaggctat ggtgagcaag ggcgaggagc tgttcaccgg
      541 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      601 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      661 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctaca acttttctat
      721 acaaagttgg caagtgcagg tgccagaaca tttctctggc ctaactggaa gacgctagcg
      781 gggggctata aaagggggtg ggggcgttcg tcctcactct agatctgcga tctaagtaag
      841 cttggcaatc cggtactgtt ggtaaagccc aactttgtat aataaagttg agctagttgc
      901 taagtctgcc gagtagaatt aaccctcact aaagggtagg tgacactatt ctaacatatc
      961 atctaacata gagggtcaac atacattcta aatcaatccc tatagtgagt cgtattagta
     1021 gacgcgagtc aatgcatgta cgacagcttt cttgtacaaa gtggtgatat cgaattcctg
     1081 cagcccgggg gatccgcccg ggctagagcg gccgccaccg cggtggagct cgcgactagt
     1141 gacgccacca tggaagatgc caaaaacatt aagaagggcc cagcgccatt ctacccactc
     1201 gaagacggga ccgccggcga gcagctgcac aaagccatga agcgctacgc cctggtgcct
     1261 ggcaccatcg cctttaccga cgcacatatc gaggtggaca ttacctacgc cgagtacttc
     1321 gagatgagcg ttcggctggc agaagctatg aagcgctatg ggctgaatac aaaccatcgg
     1381 atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg tgccctgttc
     1441 atcggtgtgg ctgtggcccc agctaacgac atctacaacg agcgcgagct gctgaacagc
     1501 atgggcatca gccagcccac cgtcgtattc gtgagcaaga aagggctgca aaagatcctc
     1561 aacgtgcaaa agaagctacc gatcatacaa aagatcatca tcatggatag caagaccgac
     1621 taccagggct tccaaagcat gtacaccttc gtgacttccc atttgccacc cggcttcaac
     1681 gagtacgact tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct gatcatgaac
     1741 agtagtggca gtaccggatt gcccaagggc gtagccctac cgcaccgcac cgcttgtgtc
     1801 cgattcagtc atgcccgcga ccccatcttc ggcaaccaga tcatccccga caccgctatc
     1861 ctcagcgtgg tgccatttca ccacggcttc ggcatgttca ccacgctggg ctacttgatc
     1921 tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg agctattctt gcgcagcttg
     1981 caagactata agattcaatc tgccctgctg gtgcccacac tatttagctt cttcgctaag
     2041 agcactctca tcgacaagta cgacctaagc aacttgcacg agatcgccag cggcggggcg
     2101 ccgctcagca aggaggtagg tgaggccgtg gccaaacgct tccacctacc aggcatccgc
     2161 cagggctacg gcctgacaga aacaaccagc gccattctga tcacccccga aggggacgac
     2221 aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt ggacttggac
     2281 accggtaaga cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg ccccatgatc
     2341 atgagcggct acgttaacaa ccccgaggct acaaacgctc tcatcgacaa ggacggctgg
     2401 ctgcacagcg gcgacatcgc ctactgggac gaggacgagc acttcttcat cgtggaccgg
     2461 ctgaagagcc tgatcaaata caagggctac caggtagccc cagccgaact ggagagcatc
     2521 ctgctgcaac accccaacat cttcgacgcc ggggtcgccg gcctgcccga cgacgatgcc
     2581 ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta aaaccataac cgagaaggag
     2641 atcgtggact atgtggccag ccaggctaca accgccaaga agctgcgcgg tggtgttgtg
     2701 ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa gatccgcgag
     2761 attctcatta aggccaagaa gggcggcaag atcgccgtgt aattcgaatt cgatatcaag
     2821 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2881 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2941 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3001 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3061 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3121 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3181 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3241 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3301 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3361 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3421 cgataccgtc gactcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3481 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3541 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3601 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3661 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcatggc
     3721 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3781 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3841 cgacgcccgg gcggcctcag tgagcgagcg agcgcgcaga gctttttgca aaagcctagg
     3901 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
     3961 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
     4021 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
     4081 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
     4141 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4201 cctaactgac acacattcca cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4261 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4321 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4381 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4441 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4501 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4561 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4621 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4681 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4741 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4801 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4861 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4921 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4981 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5041 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5101 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5161 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5221 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5281 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5341 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5401 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5461 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5521 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5581 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5641 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5701 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5761 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5821 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5881 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5941 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6001 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6061 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6121 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6181 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6241 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     6301 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     6361 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     6421 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta
     6481 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     6541 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6601 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6661 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6721 acgatgcgtc cggcgtagag gatctggcta gcgatgaccc tgctgattgg ttcgctgacc
     6781 atttccgggt gcgggacggc gttaccagaa actcagaagg ttcgtccaac caaaccgact
     6841 ctgacggcag tttacgagag agatgatagg gtctgcttca gtaagccaga tgctacacaa
     6901 ttaggcttgt acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
     6961 atacacatac gatttaggtg acactataga atacacggaa ttaattc
//
