LOCUS       sequence-259701-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hNABP2 N-terminus (1–138 aa) in E. coli with a C-terminal 
            His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Vidhyasagar V, He Y, Guo M, Ding H, Talwar T, Nguyen V, Nwosu J, 
            Katselis G, Wu Y
  TITLE     C-termini are essential and distinct for nucleic acid binding of 
            human NABP1 and NABP2.
  JOURNAL   Biochim Biophys Acta. 2016 Feb;1860(2):371-83. doi: 
            10.1016/j.bbagen.2015.11.003. Epub 2015 Nov 10.
  PUBMED    26550690
REFERENCE   2  (bases 1 to 5648)
  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"; doi: 
            "10.1016/j.bbagen.2015.11.003"; journalName: "Biochim Biophys Acta";
            date: "2016-02"; volume: "1860"; issue: "2"; pages: "371-83"
COMMENT     Sequence Label: pET29a-hNABP2-N
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     924..943
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(963..1040)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1246..1265
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1353..1372
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1353..1371
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1372..1396
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1411..1433
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1861..1878
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1931..1949)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1945..1992
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2029..2484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2116..2135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2326..2347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2576..3391)
                     /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(2576..3391)
                     /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     3299..3318
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3383..3402
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3513..4101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4002..4021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4255..4272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4287..4426
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4352..4371)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4512..4534
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4528..4719)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5528..5648,1..962))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
       61 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
      121 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
      181 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
      241 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
      301 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
      361 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
      421 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
      481 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
      541 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
      601 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
      661 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
      721 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
      781 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
      841 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
      901 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
      961 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     1021 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     1081 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     1141 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     1201 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     1261 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     1321 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     1381 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
     1441 atgacgacgg agacctttgt gaaggatatc aagcctgggc tcaagaatct gaaccttatc
     1501 ttcattgtgc tggagacagg ccgagtgacc aagacaaagg acgggcatga ggttcggacc
     1561 tgcaaagtgg cggacaaaac aggcagcatc aatatctctg tctgggacga tgttggcaat
     1621 ctgatccagc ctggggacat tatccggctc accaaagggt acgcttcagt tttcaaaggt
     1681 tgtctgacac tatgtactgg ccgtgggggt gatctgcaga agattggaga attctgtatg
     1741 gtttattctg aggttcctaa cttcagtgag ccaaacccag agtacagcac ccagcaggca
     1801 cccaacaagg cggtgcagaa cgacagcaac ccttcagctt cccagcctac cactctcgag
     1861 caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg
     1921 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1981 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     2041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2221 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2341 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2461 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2521 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
     2581 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     2641 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     2701 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     2761 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     2821 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
     2881 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     2941 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     3001 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     3061 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     3121 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     3181 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     3241 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     3301 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
     3361 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     3421 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3481 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3541 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3601 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     3661 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3721 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3781 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3841 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3901 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3961 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4021 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4081 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4141 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4201 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4261 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     4321 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4381 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4441 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4501 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4561 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4621 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4681 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4741 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4801 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4861 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4921 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4981 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5041 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5101 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     5161 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     5221 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     5281 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     5341 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     5401 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     5461 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     5521 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5581 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     5641 ccagtgag
//
