LOCUS       sequence-148002-        5264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5264)
  TITLE     pET9d-hnRNP-A1
REFERENCE   2  (bases 1 to 5264)
  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"
COMMENT     Sequence Label: pET9d-hnRNP-A1
FEATURES             Location/Qualifiers
     source          1..5264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(70..89)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     230..252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(246..437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1895..1914
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1998..2017
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1998..2016
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2048..2070
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2865..2876
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3090..3108)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3104..3151
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3517..3522,3523..3539,3540..3545))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(3558..4373)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3558..4373)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4281..4300
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4365..4384
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4984..5003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5237..5254
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
       61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
      301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
      361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
      421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
      481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
      781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
      841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
      901 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
      961 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     1021 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     1081 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     1141 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     1201 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     1261 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     1321 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     1381 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     1441 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     1501 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     1561 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     1621 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     1681 ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1741 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1801 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1861 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1921 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1981 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
     2041 aaataatttt gtttaacttt aagaaggaga tataccatgt ctaagtcaga gtctcctaaa
     2101 gagcccgaac agctgaggaa gctcttcatt ggagggttga gctttgaaac aactgatgag
     2161 agcctgagga gccattttga gcaatgggga acgctcacgg actgtgtggt aatgagagat
     2221 ccaaacacca agcgctccag gggctttggg tttgtcacat atgccactgt ggaggaggtg
     2281 gatgcagcta tgaatgcaag gccacacaag gtggatggaa gagttgtgga accaaagaga
     2341 gctgtctcca gagaagattc tcaaagacca ggtgcccact taactgtgaa aaagatattt
     2401 gttggtggca ttaaagaaga cactgaagaa catcacctaa gagattattt tgaacagtat
     2461 ggaaaaattg aagtgattga aatcatgact gaccgaggca gtggcaagaa aaggggcttt
     2521 gcctttgtaa cctttgacga ccatgactcc gtggataaga ttgtcattca gaaataccat
     2581 actgtgaatg gccacaactg tgaagttaga aaagccctgt caaagcaaga gatggctagt
     2641 gcttcatcca gccaaagagg tcgaagtggt tctggaaact ttggtggtgg tcgtggaggt
     2701 ggtttcggtg ggaatgacaa cttcggtcgt ggaggaaact tcagtggtcg tggtggcttt
     2761 ggtggcagcc gtggtggtgg tggatatggt ggcagtgggg atggctataa tggatttggt
     2821 aatgatggaa gcaattttgg aggtggtgga agctacaatg attttgggaa ttacaacaat
     2881 cagtcttcaa attttggacc catgaaggga ggaaattttg gaggcagaag ctctggcccc
     2941 tatggcggtg gaggccaata ctttgcaaaa ccacgaaacc aaggtggcta tggcggttcc
     3001 agcagcagca gtagctatgg cagtggcaga agattttaag gatccggctg ctaacaaagc
     3061 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     3121 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     3181 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
     3241 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
     3301 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
     3361 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     3421 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     3481 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     3541 gtcaaacatg agaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     3601 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     3661 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     3721 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3781 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag
     3841 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3901 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     3961 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     4021 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     4081 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     4141 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     4201 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     4261 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     4321 ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc
     4381 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaag
//
