LOCUS       sequence-252183-        5846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hNABP1 (a.k.a hSSB2 or SOSSB2) 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 5846)
  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 5846)
  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
FEATURES             Location/Qualifiers
     source          1..5846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..104
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     192..211
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        192..210
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    211..235
                     /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             250..272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             898..915
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(968..986)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      982..1029
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1066..1521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1153..1172)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1363..1384
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1613..2428)
                     /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(1613..2428)
                     /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     2336..2355
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2420..2439
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2550..3138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3039..3058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3292..3309
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3324..3463
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3389..3408)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3549..3571
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3565..3756)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4565..5647)
                     /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     5609..5628
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5648..5725)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
       61 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
      121 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga
      181 tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct
      241 agaaataatt ttgtttaact ttaagaagga gatatacata tgaatagggt caacgaccca
      301 cttattttta taagagatat taagcccgga ctgaaaaact taaatgtcgt ctttattgtc
      361 ctggagatag gacgcgtgac caaaaccaaa gacggccatg aagtgagatc gtgcaaagta
      421 gcagataaaa cgggcagcat cactatttcc gtgtgggatg agatcggagg tcttatacag
      481 ccaggggata ttattcggtt gaccagaggg tatgcatcca tgtggaaagg atgtctgaca
      541 ctttatactg gaaggggtgg tgaacttcaa aaaattgggg aattttgtat ggtttattca
      601 gaagtgccaa atttcagtga acccaaccca gattatcgag gacagcagaa caaaggggca
      661 cagagtgaac agaagaataa ttccatgaat agtaatatgg gtacaggtac atttggacca
      721 gtgggaaatg gtgttcacac tggccctgaa tcaagggaac accagttttc acatgctggc
      781 agaagcaatg gccggggact tataaatcca caactacaag gaacagctag taatcaaaca
      841 gtgatgacca caataagtaa tggcagggac cctcggagag cctttaaaag actcgagcac
      901 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      961 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     1021 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     1081 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1141 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1201 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1261 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1321 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1381 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1441 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1501 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1561 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     1621 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1681 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1741 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     1801 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     1861 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     1921 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     1981 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2041 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2101 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2161 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2221 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2281 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2341 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     2401 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     2461 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     2701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     2941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3241 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3301 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3361 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3421 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3481 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3541 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3601 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3661 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3721 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3781 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3841 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3901 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3961 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     4021 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     4081 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4141 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     4201 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     4261 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4321 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4381 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4441 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     4501 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     4681 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     4741 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     4801 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc
     4861 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     4921 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     4981 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     5041 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     5101 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     5161 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     5221 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     5281 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     5341 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     5401 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     5461 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     5521 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     5581 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     5641 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     5701 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     5761 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     5821 gcatgcaagg agatggcgcc caacag
//
