LOCUS       sequence-310519-        6660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding the translational pair that enables efficient 
            incorporation of Benzoyl-phenylalanine in response to the amber 
            codon into proteins in mammmalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6660)
  AUTHORS   Bottke T, Ernicke S, Serfling R, Ihling C, Burda E, Gurevich VV, 
            Sinz A, Coin I
  TITLE     Exploring GPCR-arrestin interfaces with genetically encoded 
            crosslinkers.
  JOURNAL   EMBO Rep. 2020 Sep 14:e50437. doi: 10.15252/embr.202050437.
  PUBMED    32929862
REFERENCE   2  (bases 1 to 6660)
  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: "EMBO Rep. 
            2020 Sep 14:e50437. doi: 10.15252/embr.202050437."
COMMENT     Sequence Label: pIRE4-Bpa
FEATURES             Location/Qualifiers
     source          1..6660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..500
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(36..59)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     433..452
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      527..536
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1958..2079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1995..2014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2049..2068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2104..2352
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        2462..2710
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        2820..3068
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        3178..3426
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     rep_origin      complement(3529..3984)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3666..3687)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3878..3897
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4011..4115
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4140..4160)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4256..4466
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4278..4474
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4325..4460
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4509..5303
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4509..5303
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4563..4582)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5173..5192
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5491..5510)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5535..5582
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5933..6521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6422..6441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
       61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
      121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
      181 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
      241 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
      301 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
      361 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
      421 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
      481 cctcatctcc gggcctttcg acctgctagc gtttaaactt aagcttgcca ccatggccag
      541 cagcaacctg atcaagcagc tgcaggaacg gggcctggtg gcccaagtga cagatgaaga
      601 agccctggcc gagagactgg cccagggacc tattgccctg ggctgcggct ttgaccccac
      661 cgccgattct ctgcacctgg gacatctggt gcctctgctg tgtctgaagc ggttccagca
      721 ggccggccac aaacctgtgg ctctcgtggg cggagccacc ggcctgattg gcgatcctag
      781 ctttaaggcc gccgagcgga agctgaacac cgaagagaca gtgcaggaat gggtggacaa
      841 gatccggaaa caggtggccc ccttcctgga cttcgactgc ggcgagaata gcgccattgc
      901 cgccaacaac tacgattggt tcggcaatat gaacgtgctg accttcctgc gggacatcgg
      961 caagcacttc agcgtgaacc agatgatcaa caaagaagcc gtgaagcagc ggctgaacag
     1021 agaggaccag ggcatcagct tcaccgagtt cagctacaac ctgctgcagg gctacggctt
     1081 cgcctgcgcc aacaagcagt acggcgtggt gctgcagatc ggcggctctg accagtgggg
     1141 caatatcacc agcggcatcg acctgaccag acggctgcac cagaaccagg tgttcggcct
     1201 gaccgtgccc ctgatcacaa aggccgacgg caccaagttc ggcaagaccg aaggcggagc
     1261 cgtgtggctg gaccccaaga aaaccagccc ctacaagttc taccagttct ggatcaacac
     1321 cgccagagcc gacgtgtacc ggttcctgaa gttcttcacc ttcatgagca tcgaggaaat
     1381 caacgccctg gaagaagagg acaagaacag cggcaaggcc cccagagccc agtacgtgct
     1441 ggcagaacaa gtgaccaggc tggtgcacgg cgaggaagga ctgcaggccg ccaagagaat
     1501 caccgagtgc ctgttcagcg gcagcctgag cgctctgagc gaggccgatt ttgagcagct
     1561 ggctcaggac ggcgtgccca tggtggaaat ggaaaagggc gccgacctga tgcaggccct
     1621 ggtggattct gagctgcagc ccagcagagg acaggccaga aagacaatcg ccagcaacgc
     1681 catcaccatc aacggcgaga agcagagcga ccccgagtat ttcttcaaag aggaagatcg
     1741 gctgttcggc cggttcaccc tgctgcggag aggcaagaag aactactgcc tgatctgctg
     1801 gaagtaataa gaattctgca gatatccagc acagtggcgg ccgcgactct agatcataat
     1861 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1921 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1981 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2041 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggtctagact cgaggggcag
     2101 gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     2161 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     2221 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     2281 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     2341 ggacgaaaca ccggaggggt agcgaagtgg ctaaacgcgg cggactctaa atccgctccc
     2401 tttgggttcg gcggttcgaa tccgtccccc tccagtcctt tttttggtcg aggggcagga
     2461 agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     2521 gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     2581 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     2641 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     2701 acgaaacacc ggaggggtag cgaagtggct aaacgcggcg gactctaaat ccgctccctt
     2761 tgggttcggc ggttcgaatc cgtccccctc cagtcctttt tttggtcgag gggcaggaag
     2821 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     2881 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     2941 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     3001 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     3061 gaaacaccgg aggggtagcg aagtggctaa acgcggcgga ctctaaatcc gctccctttg
     3121 ggttcggcgg ttcgaatccg tccccctcca gtcctttttt tggtcgaggg gcaggaagag
     3181 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     3241 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     3301 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     3361 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     3421 aacaccggag gggtagcgaa gtggctaaac gcggcggact ctaaatccgc tccctttggg
     3481 ttcggcggtt cgaatccgtc cccctccagt cctttttttg gtcgacgtaa attgtaagcg
     3541 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3601 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3661 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3721 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3781 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3841 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3901 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3961 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4081 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4141 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4201 aagcatgcat ctcaattagt cagcaaccak rkkyssrmmg tccccaggct ccccagcagg
     4261 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4321 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4381 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4441 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4501 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4561 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4621 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4681 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4741 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4801 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4861 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4921 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4981 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     5041 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5101 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5161 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5221 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5281 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5341 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5401 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5461 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5521 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5581 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5641 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5701 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag tactaaatat
     5761 ttatcccggg ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5821 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5881 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5941 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6001 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6061 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6121 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6181 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6241 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6301 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6361 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6421 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6481 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6541 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6601 ctgattctgt ggataaccgt attaccgcca tgcatacgcg ttagttatta aaattctacc
//
