LOCUS       sequence-252189-        5480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hNABP1 C-terminus (125–204 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 5480)
  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 5480)
  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-hNABP1-C
FEATURES             Location/Qualifiers
     source          1..5480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..58
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        39..57
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    58..82
                     /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             97..119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             379..396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(449..467)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      463..510
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      547..1002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(634..653)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     844..865
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1094..1909)
                     /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(1094..1909)
                     /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     1817..1836
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1901..1920
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2031..2619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2520..2539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2773..2790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2805..2944
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2870..2889)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3030..3052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3046..3237)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4046..5128)
                     /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"
     primer_bind     5090..5109
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5129..5206)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5412..5431
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca ctatagggga
       61 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      121 atacatatga acaaaggggc acagagtgaa cagaagaata attccatgaa tagtaatatg
      181 ggtacaggta catttggacc agtgggaaat ggtgttcaca ctggccctga atcaagggaa
      241 caccagtttt cacatgctgg cagaagcaat ggccggggac ttataaatcc acaactacaa
      301 ggaacagcta gtaatcaaac agtgatgacc acaataagta atggcaggga ccctcggaga
      361 gcctttaaaa gactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
      421 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
      481 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
      541 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      601 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      661 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      721 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      781 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      841 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      901 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      961 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1081 catgaattaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1141 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1201 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1261 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1321 catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt
     1381 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     1441 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     1501 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     1561 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     1621 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     1681 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     1741 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     1801 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     1861 aatttaatcg cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg
     1921 tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag
     1981 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2041 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2101 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2161 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2221 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2281 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2341 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2401 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2461 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2521 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2581 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2641 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2701 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2761 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2821 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
     2881 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     2941 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3001 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3061 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     3121 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     3181 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     3241 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     3301 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
     3361 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
     3421 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
     3481 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
     3541 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
     3601 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
     3661 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
     3721 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     3781 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     3841 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     3901 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     3961 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     4021 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4081 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     4141 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     4201 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4261 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4321 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4381 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4441 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4501 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     4561 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     4621 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     4681 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     4741 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     4801 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     4861 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     4921 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     4981 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     5041 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     5101 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     5161 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     5221 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     5281 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     5341 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     5401 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5461 atgccggcca cgatgcgtcc
//
