LOCUS       sequence-385804-       10399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ALFA-tagged GluRIIA-Construct for integration at attP sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10399)
  AUTHORS   Beckers CJ, Mrestani A, Komma F, Dannhauser S
  TITLE     Versatile Endogenous Editing of GluRIIA in Drosophila melanogaster.
  JOURNAL   Cells. 2024 Feb 10;13(4):323. doi: 10.3390/cells13040323.
  PUBMED    38391936
REFERENCE   2  (bases 1 to 10399)
  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: "Cells."; 
            date: "2024-02-10"; volume: "13(4):323. doi"; pages: " 
            10.3390/cells13040323"
COMMENT     Sequence Label: pCB20
FEATURES             Location/Qualifiers
     source          1..10399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..31,32..233)
                     /label=P element 3' end
                     /note="P element 3' end"
     protein_bind    353..386
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             4047..4085
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     protein_bind    4691..4724
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(7772..8326,8327..8357)
                     /label=P element 5' end
     rep_origin      complement(8592..9180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8672..8691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9351..10142,10143..10211))
                     /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     9974..9993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10212..10316)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgatgaaa taacataagg tggtcccgtc ggcaagagac atccacttaa cgtatgcttg
       61 caataagtgc gagtgaaagg aatagtattc tgagtgtcgt attgagtctg agtgagacag
      121 cgatatgatt gttgattaac ccttagcatg tccgtggggt ttgaattaac tcataatatt
      181 aattagacga aattattttt aaagttttat ttttaataat ttgcgagtac gcaaagctag
      241 agttgttggt tggcacacca caaatatact gttgccgagc acaattccgc tcgaattgcc
      301 ggctgtacac ttaagaagtt cctactgcag gtaatgaatt gccgcggtgc gggtgccagg
      361 gcgtgccctt gggctccccg ggcgcgtact ccacgaattc actggagaga gcttcccatc
      421 ggctgagatg acattcttgg ccgtgccagt gagttccggc agtcagtagg cgtgcgaagc
      481 gctccaagac tgggtgcttc cgttgcgtga aaataaattg tgaagaagaa agacggcact
      541 ttgtatctgc tcgttttgaa tttttttttt ctgtggcagt ctgtgtgcgt ctgtagatat
      601 agatttgtac atatgcagtt gggaagtgaa cacgaaatgg ctgctgcatt tagaaaattc
      661 aattagtttc gagtgtgttg aggtaaacga aaaaatgcgc ctttgtccag ttgtcatcta
      721 tgcatttata atcatcatcg ggtttctgga ggggattata gcccttggtg gcgacgatcg
      781 caatgaaatc acagtgggta ggtccctcca ttaattactt agtgatagca atagctcaac
      841 gttttgccct cgaaggtgcc attttctacg agaacgagaa ggagatcgag ctcagcttcg
      901 atcaggcctt tcgcgaggtc aacaatatga agttctcgga gctgcgattc gtgaccatca
      961 aacgctatat gcccaccaac gacagtttcc ttttgcagca gattagtaag taaatacgct
     1021 cggatgcggc agaaccaagt tacgagaaat tcatcttccg tgtgtagcat gtgaactgat
     1081 ttccaatggc gttgcggcta ttttcggacc aagttcaaag gcagccagtg gtaagaaaag
     1141 cacacacaca cacacacact gtgtgtggac atggcccaaa acatatgttc gcgatgtgac
     1201 ggttcccccc tttcgtggca gatattgttg cgcaaatcgc gaacgcgacc ggcattccac
     1261 acattgagta cgatttgaag ctggaggcca cgcgtcagga gcagctcaac caccagatgt
     1321 ccatcaatgt ggcaccctcc ttgtcagtac tctcgcgtgc ctacttcgag atcatcaaat
     1381 ccaactatga gtggcgcacc ttcactctga tctatgagac tccggagggt aagtagtgcc
     1441 tggatacccc gtagagatat ggactacatt tgaagtcctt aggcctggcg cgcctgcagg
     1501 atctgatgaa catccaagcg ctgaacagtg actatgtcaa gctgcgaaat ttggcagact
     1561 atgcggatga ctatcggata ttgtggaagg agacagacga gactttccac gagcagcgca
     1621 tcattttgga ctgtgagccg aagaccctta aggagctgct taaagtatcc atagacttta
     1681 agttgcaggg cccgtttagg gtaattcttc atttatgaaa gcgagaaagt acttactaag
     1741 tcgctttatc tttagaactg gtttctcacc catctggata cgcataactc gggtctgagg
     1801 gatatttata atgaggactt caaggccaac attacgtcgg tgcgattgaa agtggtcgac
     1861 gcgaatccat tcgagagaaa gaagacccgg ctcaccaaag tggatgtaag tggaggccca
     1921 actgactgta accttggatc taatctagcg tacttcgcag caaatattgg ggaaccaaac
     1981 catgttgccc attctcatct acgacgctgt cgtcctgttt gccagttctg cccgaaatgt
     2041 tattgctgcc atgcagccct ttcatccacc caatcgccat tgcggcagtt cgtctccttg
     2101 gatgctggga gcatttatag tcaacgagat gaagaccatt tccgaggatg atgtggaacc
     2161 gcactttaag accgaaaata tgaagcttga cgagtatggg cagcgcatcc acttcaacct
     2221 ggagatctac aagccaacgg tgaacgaacc aatgatggtc tggacaccag acaatgggat
     2281 aaagaagcgt ctcctcaact tggaactgga aagtgccggc accacccagg atttctcgga
     2341 gcagcgtaaa gtgtatacag tggttaccca ctacgaggag ccctacttca tgatgaagtt
     2401 ggttttgaag cagtaatctg gtttttagca gactacaacc tttccttatt tcctttaggg
     2461 aggatcacga gaacttcaga ggcagggaaa agtacgaggg ctatgccgtg gatctcatta
     2521 gcaagctttc cgagctcatg gagttcgatt acgagttcat gatcgttaat ggcaatggca
     2581 aatacaatcc ggagacgaag caatgggatg gcattatacg gaaactgata gaccacgtag
     2641 gtttcaccag aagtatagta gtaactccac ttgatatttg tatcttttat acacagcatg
     2701 cccaaatcgg cgtctgcgat ctgaccatca cccaaatgcg acgttctgtg gtggacttca
     2761 cggtgccctt catgcagttg ggcattagca ttctgcacta caagagtcct ccggagccga
     2821 agaatcagtt tgccttcttg gaaccgtttg ccgtcgaagt ctggatctat atgatctttg
     2881 cccagttgat aatgactttg gcttttgttt tcatagccag gtgggattag gttaacgctc
     2941 ttgagtgaag cttaatttac ttttcttaca gactttcgta tcgcgaatgg ttgccaccta
     3001 atcccgccat tcaggatcca gatgagctgg aaaatatctg gaatgtgaac aactcgactt
     3061 ggctgatggt gggctccatc atgcaacagg gctgcgacat tctgcctagg tctcagatta
     3121 tatattacac agtatcaatg ggtatcgaaa tgtaaacttt cttcactcac agaggtccac
     3181 acatgcgcat actcactggc atgtggtggt tcttcgccct gatgatgctc agtacctaca
     3241 cggccaactt ggccgccttc ctgaccagca acaagtggca gagcagcata aagagcctcc
     3301 aggacctcat cgaacaggat aaggtgcact tcggcagcat gcgcggtggc agcacctcgt
     3361 tgttcttttc ggagtcgaat gacacggatt accagcgtgc gtggaaccag atgaaggact
     3421 tcaatccctc ggccttcacc agcaccaaca aagagggagt ggcacgtgtg cgcaaggaga
     3481 agggcggcta tgcctttctg atggagacca ccagtttgac ctacaatatc gaacggaact
     3541 gcgatctcac ccagattggc gagcagatcg gggagaagca ctacggcctg gccgtgccac
     3601 tgggtgagta gctgtgccgc cagtggaggg agttgggata ttactttctt ccatttcaac
     3661 caggctctga ttaccggacc aacctcagcg tgtccattct ccagctgagc gaaaggggcg
     3721 agctgcagaa gatgaagaac aagtggtgga agaaccacaa tgtcacctgt gactcctacc
     3781 acgaggtgga tggtgacgag ctgtccatta ttgagctggg cggcgtgttt cttgttctgg
     3841 ccggtggcgt tctcatcggc gttatcctgg gcatctttga gttcctgtgg aacgtgcaga
     3901 acgtggcggt ggaggagcga gtgactccgt ggcaggcttt caaggcggag ctcatctttg
     3961 ccctgaagtt ttgggtgcgc aaaaagccga tgcgcatctc cagcagcagc gacaagtcgt
     4021 cgtccaggcg atcatccggt tcccccagcc gcctggagga ggagctgcgc cgccgcctga
     4081 ccgagcccag acgcagctcc aaggaaaagt cccgcagcaa gacggttagc taggtggtcg
     4141 gaatattgga cgattggcgt cttctacttt ggcatttcat tagagagtcg cattggaaat
     4201 gaggcactcc atggtgtcca cattgtatat tgtgagctat atttgtagaa ttattttaga
     4261 acacagatag cgcatttggg tatattgaat gttctgaaga tttaacgaaa ctttcaatga
     4321 aataatttgg ataactaatt ttagtaaact tttctgttgg ctctaatgtt ttaatagttg
     4381 acaataattg attttcctta caaagattta ataaataaac attaaaatat aatatgtgtg
     4441 tttatataca atatatagtc agttagttgt gaaattatat tctatacatt tcttttgaaa
     4501 gctgcctaat tttagaccac atttagtttt cattcactgg tccatttcca gctaatgctc
     4561 gtgtctgcat ggtctagatt aaagagtata gagtatagac gctagactct aaagtctaga
     4621 catgtggtga ttgctcttct cgccgagcat ggaatccatc cacaatggct gctgttctcc
     4681 atgaggtacc ataacttcgt ataatgtatg ctatacgaag ttatcgtacg actagcacct
     4741 gactgtcgag cccagtggaa acccttgaaa tgcctttaag tcgagcccag tggaaaccct
     4801 tgaaatgcct ttaagtcgag cccagtggaa acccttgaaa tgcctttaag tcgagcccag
     4861 tggaaaccct tgaaatgcct ttaagtcgag cccagtggaa acccttgaaa tgcctttaag
     4921 tcgagtagca gtcgaggtag gtacgccatg ccccgttatt ctctattcgt tttgtgactc
     4981 tccctctctg tactattgct ctctcactct gtcgcacagt aaacggcacg ctattctcgt
     5041 tgcttcgaga gagcgcgcct cgaatgttcg cgaaaagagc gccggagtat aaatagaggc
     5101 gcttcgtcga cggagcgtca attcaattca aacaagcaaa gtgaacacat cgcgaagcgt
     5161 aagctgagca aacaaacaag cgcagctgaa caagctaaac aatctgcaat aaagtgcaag
     5221 ttaaagtgaa tcaattaaaa gtaaccaaca accaagtaat taaactaaaa actgcaacta
     5281 ctgaaatcaa ccaagaagtc attattgaag acaagaagag aactctgaat aggtcgatag
     5341 cgtcaattcg gcacgagcgg tttcgtgacg aagctccaag cggtttacgc catcaattaa
     5401 acacaaagtg ctgtgccaaa actcctctcg cttcttattt ttgtttgttt tttgagtgat
     5461 tgggtgggtg attggttttg ggtgggtaag caggggaaag tgtgaaaaat cccggcaatg
     5521 ggccaagagg atcaggagct attaattcgc ggaggcagca aacacccatc tgccgagcat
     5581 ctgaacaatg gtgacagcgg agcggcttcg cagagctgca ttaaccaggg cttcgggcag
     5641 gccaaaaact acggcacgct cctgccaccc agtccgccgg aggactccgg ttcagggagc
     5701 ggccaactag ccgagaacct cacctatgcc tggcacaata tggacatctt tggggcggtc
     5761 aatcagccgg gctccggatg gcggcagctg gtcaaccgga cacgcggact attctgcaac
     5821 gagcgacaca taccggcgcc caggaaacat ttgctcaaga acgtttgcgg cgtggcctat
     5881 ccgggcgaac ttttggccgt gatgggcagt tccggtgccg gaaagacgac cctgctgaat
     5941 gcccttgcct ttcgatcgcc gcagggcatc caagtatcgc catccgggat gcgactgctc
     6001 aatggccaac ctgtggacgc caaggagatg caggccaggt gcgcctatgt ccagcaggat
     6061 gacctcttta tcggctccct aacggccagg gaacacctga ttttccaggc catggtgcgg
     6121 atgccacgac atctgaccta tcggcagcga gtggcccgcg tggatcaggt gatccaggag
     6181 ctttcgctca gcaaatgtca gcacacgatc atcggtgtgc ccggcagggt gaaaggtctg
     6241 tccggcggag aaaggaagcg tctggcattc gcctccgagg cactaaccga tccgccgctt
     6301 ctgatctgcg atgagcccac ctccggactg gactcattta ccgcccacag cgtcgtccag
     6361 gtgctgaaga agctgtcgca gaagggcaag accgtcatcc tgaccattca tcagccgtct
     6421 tccgagctgt ttgagctctt tgacaagatc cttctgatgg ccgagggcag ggtagctttc
     6481 ttgggcactc ccagcgaagc cgtcgacttc ttttcctacg tgggtgccca gtgtcctacc
     6541 aactacaatc cggcggactt ttacgtacag gtgttggccg ttgtgcccgg acgggagatc
     6601 gagtcccgtg atcggatcgc caagatatgc gacaattttg ccattagcaa agtagcccgg
     6661 gatatggagc agttgttggc caccaaaaat ttggagaagc cactggagca gccggagaat
     6721 gggtacacct acaaggccac ctggttcatg cagttccggg cggtcctgtg gcgatcctgg
     6781 ctgtcggtgc tcaaggaacc actcctcgta aaagtgcgac ttattcagac aacgatggtt
     6841 gccatcttga ttggcctcat ctttttgggc caacaactca cgcaagtggg cgtgatgaat
     6901 atcaacggag ccatcttcct cttcctgacc aacatgacct ttcaaaacgt ctttgccacg
     6961 ataaatgtgt tcacctcaga gctgccagtt tttatgaggg aggcccgaag tcgactttat
     7021 cgctgtgaca catactttct gggcaaaacg attgccgaat tgccgctttt tctcacagtg
     7081 ccactggtct tcacggcgat tgcctatccg atgatcggac tgcgggccgg agtgctgcac
     7141 ttcttcaact gcctggcgct ggtcactctg gtggccaatg tgtcaacgtc cttcggatat
     7201 ctaatatcct gcgccagctc ctcgacctcg atggcgctgt ctgtgggtcc gccggttatc
     7261 ataccattcc tgctctttgg cggcttcttc ttgaactcgg gctcggtgcc agtatacctc
     7321 aaatggttgt cgtacctctc atggttccgt tacgccaacg agggtctgct gattaaccaa
     7381 tgggcggacg tggagccggg cgaaattagc tgcacatcgt cgaacaccac gtgccccagt
     7441 tcgggcaagg tcatcctgga gacgcttaac ttctccgccg ccgatctgcc gctggactac
     7501 gtgggtctgg ccattctcat cgtgagcttc cgggtgctcg catatctggc tctaagactt
     7561 cgggcccgac gcaaggagta gccgacatat atccgaaata actgtttgtt tttttttttt
     7621 accattatta ccatcgtgtt tactgtttat tgccccctca aaaagctaat gtaattatat
     7681 ttgtgccaat aaaaacaaga tatgacctat agaatacaaa aaaaaaaaaa aaaaaaaaaa
     7741 aaaaactcga caccgggatc caaacctagt ggctgctgct ctaaacgacg catttcgtac
     7801 tccaaagtac gaattttttc cctcaagctc ttattttcat taaacaatga acaggaccta
     7861 acgcacagtc acgttattgt ttacataaat gatttttttt actattcaaa cttactctgt
     7921 ttgtgtactc ccactggtat agccttcttt tatcttttct ggttcaggct ctatcacttt
     7981 actaggtacg gcatctgcgt tgagtcgcct ccttttaaat gtctgacctt ttgcaggtgc
     8041 agccttccac tgcgaatcat taaagtgggt atcacaaatt tgggagtttt caccaaggct
     8101 gcacccaagg ctctgctccc acaattttct cttaatagca cacttcggca cgtgaattaa
     8161 ttttactcca gtcacagctt tgcagcaaaa tttgcaatat ttcatttttt tttattccac
     8221 gtaagggtta atgttttcaa aaaaaaattc gtccgcacac aacctttcct ctcaacaagc
     8281 aaacgtgcac tgaatttaag tgtatacttc ggtaagcttc ggctatcgac gggaccacct
     8341 tatgttattt catcatgggc cagacccacg tagtccagcg gcagatcggc ggcggagaag
     8401 ttaagcgtct ccaggatgac cttgcccgaa ctggggcacg tggtgttcga cgatgtgcag
     8461 ctaatttcgc ccggctccac gtccgcccat tggttaatca gcagaccctc gttggcgtaa
     8521 cggaaccatg agaggtacga caaccatttg aggtatactg gcaccgagcc cgagttcaag
     8581 aagaaggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     9001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     9061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     9121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     9181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     9241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9301 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     9901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     9961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10321 tttccccgaa aagtgccacc tcgaagtctt agagccagat atgcgagcac ccggaagctc
    10381 acgatgagaa tggccagac
//
