LOCUS       sequence-299690-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For recombination of HDLBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  AUTHORS   Zinnall U, Milek M, Minia I, Vieira-Vieira CH, Muller S, Mastrobuoni
            G, Hazapis OG, Del Giudice S, Schwefel D, Bley N, Voigt F, Chao JA, 
            Kempa S, Huttelmaier S, Selbach M, Landthaler M
  TITLE     HDLBP binds ER-targeted mRNAs by multivalent interactions to promote
            protein synthesis of transmembrane and secreted proteins.
  JOURNAL   Nat Commun. 2022 May 18;13(1):2727. doi: 10.1038/s41467-022-30322-7.
  PUBMED    35585045
REFERENCE   2  (bases 1 to 6357)
  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: "Nat 
            Commun."; date: "2022-05-18"; volume: "13(1):2727. doi"; pages: " 
            10.1038/s41467-022-30322-7"
COMMENT     Sequence Label: pDONR221_HDLBP
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4482..4581)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4598..4617)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4599..4617)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4622..4638)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4622..4638)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4740..4759)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4751..5560
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4818..4837)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5707..6295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6196..6215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatgagtt ccgttgcagt tttgacccaa gagagttttg ctgaacaccg
      721 aagtgggctg gttccgcaac aaatcaaagt tgccactcta aattcagaag aggagagcga
      781 ccctccaacc tacaaggatg ccttccctcc acttcctgag aaagctgctt gcctggaaag
      841 tgcccaggaa cccgctggag cctgggggaa caagatccga cccatcaagg cttctgtcat
      901 cactcaggtg ttccatgtac ccctggagga gagaaaatac aaggatatga accagtttgg
      961 agaaggtgaa caagcaaaaa tctgccttga gatcatgcag agaactggtg ctcacttgga
     1021 gctgtctttg gccaaagacc aaggcctctc catcatggtg tcaggaaagc tggatgctgt
     1081 catgaaagct cggaaggaca ttgttgctag actgcagact caggcctcag caactgttgc
     1141 cattcccaaa gaacaccatc gctttgttat tggcaaaaat ggagagaaac tgcaagactt
     1201 ggagctaaaa actgcaacca aaatccagat cccacgccca gatgacccca gcaatcagat
     1261 caagatcact ggcaccaaag agggcatcga gaaagctcgc catgaagtct tactcatctc
     1321 tgccgagcag gacaaacgtg ctgtggagag gctagaagta gaaaaggcat tccacccctt
     1381 catcgctggg ccgtataata gactggttgg cgagatcatg caggagacag gcacgcgcat
     1441 caacatcccc ccacccagcg tgaaccggac agagattgtc ttcactggag agaaggaaca
     1501 gttggctcag gctgtggctc gcatcaagaa gatttatgag gagaagaaaa agaagactac
     1561 aaccattgca gtggaagtga agaaatccca acacaagtat gtcattgggc ccaagggcaa
     1621 ttcattgcag gagatccttg agagaactgg agtttccgtt gagatcccac cctcagacag
     1681 catctctgag actgtaatac ttcgaggcga acctgaaaag ttaggtcagg cgttgactga
     1741 agtctatgcc aaggccaata gcttcaccgt ctcctctgtc gccgcccctt cctggcttca
     1801 ccgtttcatc attggcaaga aagggcagaa cctggccaaa atcactcagc agatgccaaa
     1861 ggttcacatc gagttcacag agggcgaaga caagatcacc ctggagggcc ctacagagga
     1921 tgtcaatgtg gcccaggaac agatagaagg catggtcaaa gatttgatta accggatgga
     1981 ctatgtggag atcaacatcg accacaagtt ccacaggcac ctcattggga agagcggtgc
     2041 caacataaac agaatcaaag accagtacaa ggtgtccgtg cgcatccctc ctgacagtga
     2101 gaagagcaat ttgatccgca tcgaggggga cccacagggc gtgcagcagg ccaagcgaga
     2161 gctgctggag cttgcatctc gcatggaaaa tgagcgtacc aaggatctaa tcattgagca
     2221 aagatttcat cgcacaatca ttgggcagaa gggtgaacgg atccgtgaaa ttcgtgacaa
     2281 attcccagag gtcatcatta actttccaga cccagcacaa aaaagtgaca ttgtccagct
     2341 cagaggacct aagaatgagg tggaaaaatg cacaaaatac atgcagaaga tggtggcaga
     2401 tctggtggaa aatagctatt caatttctgt tccgatcttc aaacagtttc acaagaatat
     2461 cattgggaaa ggaggcgcaa acattaaaaa gattcgtgaa gaaagcaaca ccaaaatcga
     2521 ccttccagca gagaatagca attcagagac cattatcatc acaggcaagc gagccaactg
     2581 cgaagctgcc cggagcagga ttctgtctat tcagaaagac ctggccaaca tagccgaggt
     2641 agaggtctcc atccctgcca agctgcacaa ctccctcatt ggcaccaagg gccgtctgat
     2701 ccgctccatc atggaggagt gcggcggggt ccacattcac tttcccgtgg aaggttcagg
     2761 aagcgacacc gttgttatca ggggcccttc ctcggatgtg gagaaggcca agaagcagct
     2821 cctgcatctg gcggaggaga agcaaaccaa gagtttcact gttgacatcc gcgccaagcc
     2881 agaataccac aaattcctca tcggcaaggg gggcggcaaa attcgcaagg tgcgcgacag
     2941 cactggagca cgtgtcatct tccctgcggc tgaggacaag gaccaggacc tgatcaccat
     3001 cattggaaag gaggacgccg tccgagaggc acagaaggag ctggaggcct tgatccaaaa
     3061 cctggataat gtggtggaag actccatgct ggtggacccc aagcaccacc gccacttcgt
     3121 catccgcaga ggccaggtct tgcgggagat tgctgaagag tatggcgggg tgatggtcag
     3181 cttcccacgc tctggcacac agagcgacaa agtcaccctc aagggcgcca aggactgtgt
     3241 ggaggcagcc aagaaacgca ttcaggagat cattgaggac ctggaagctc aggtgacatt
     3301 agaatgtgct ataccccaga aattccatcg atctgtcatg ggccccaaag gttccagaat
     3361 ccagcagatt actcgggatt tcagtgttca aattaaattc ccagacagag aggagaacgc
     3421 agttcacagt acagagccag ttgtccagga gaatggggac gaagctgggg aggggagaga
     3481 ggctaaagat tgtgaccccg gctctccaag gaggtgtgac atcatcatca tctctggccg
     3541 gaaagaaaag tgtgaggctg ccaaggaagc tctggaggca ttggttcctg tcaccattga
     3601 agtagaggtg ccctttgacc ttcaccgtta cgttattggg cagaaaggaa gtgggatccg
     3661 caagatgatg gatgagtttg aggtgaacat acatgtcccg gcacctgagc tgcagtctga
     3721 catcatcgcc atcacgggcc tcgctgcaaa tttggaccgg gccaaggctg gactgctgga
     3781 gcgtgtgaag gagctacagg ccgagcagga ggaccgggct ttaaggagtt ttaagctgag
     3841 tgtcactgta gaccccaaat accatcccaa gattatcggg agaaaggggg cagtaattac
     3901 ccaaatccgg ttggagcatg acgtgaacat ccagtttcct gataaggacg atgggaacca
     3961 gccccaggac caaattacca tcacagggta cgaaaagaac acagaagctg ccagggatgc
     4021 tatactgaga attgtgggtg aacttgagca gatggtttct gaggacgtcc cgctggacca
     4081 ccgcgttcac gcccgcatca ttggtgcccg cggcaaagcc attcgcaaaa tcatggacga
     4141 attcaaggtg gacattcgct tcccacagag cggagcccca gaccccaact gcgtcactgt
     4201 gacggggctc ccagagaatg tggaggaagc catcgaccac atcctcaatc tggaggagga
     4261 atacctagct gacgtggtgg acagtgaggc gctgcaggta tacatgaaac ccccagcaca
     4321 cgaagaggcc aaggcacctt ccagaggctt tgtggtgcgg gacgcaccct ggaccgccag
     4381 cagcagtgag aaggctcctg acatgagcag ctctgaggaa tttcccagct ttggggctca
     4441 ggtggctccc aagaccctcc cttggggccc caaacgatag gacccagctt tcttgtacaa
     4501 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     4561 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     4621 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     4681 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     4741 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     4801 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4861 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4921 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4981 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     5041 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     5101 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     5161 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     5221 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     5281 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     5341 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     5401 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     5461 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     5521 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     5581 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     5641 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5701 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5761 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5821 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5881 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5941 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6001 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6061 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     6121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
