LOCUS       sequence-252184-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express hNABP2 (a.k.a hSSB1 or SOSSB1) 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 5867)
  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 5867)
  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
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(42..1124)
                     /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     1086..1105
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1125..1202)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1408..1427
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1515..1534
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1515..1533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1534..1558
                     /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             1573..1595
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2242..2259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2312..2330)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2326..2373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2410..2865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2497..2516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2707..2728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2957..3772)
                     /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(2957..3772)
                     /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     3680..3699
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3764..3783
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3894..4482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4383..4402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4636..4653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4668..4807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4733..4752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4893..4915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4909..5100)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
       61 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      121 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
      181 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
      241 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
      301 tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc ggtaatggcg
      361 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
      421 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
      481 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
      541 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
      601 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
      661 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
      721 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
      781 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
      841 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
      901 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
      961 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     1021 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     1081 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     1141 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     1201 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     1261 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     1321 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     1381 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     1441 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatcgatct
     1501 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     1561 tctagaaata attttgttta actttaagaa ggagatatac atatgacgac ggagaccttt
     1621 gtgaaggata tcaagcctgg gctcaagaat ctgaacctta tcttcattgt gctggagaca
     1681 ggccgagtga ccaagacaaa ggacgggcat gaggttcgga cctgcaaagt ggcggacaaa
     1741 acaggcagca tcaatatctc tgtctgggac gatgttggca atctgatcca gcctggggac
     1801 attatccggc tcaccaaagg gtacgcttca gttttcaaag gttgtctgac actatgtact
     1861 ggccgtgggg gtgatctgca gaagattgga gaattctgta tggtttattc tgaggttcct
     1921 aacttcagtg agccaaaccc agagtacagc acccagcagg cacccaacaa ggcggtgcag
     1981 aacgacagca acccttcagc ttcccagcct accactggac cctctgctgc ctctccagcc
     2041 tctgagaacc agaatgggaa tggactgagt gccccaccag gtcccggtgg tggcccacat
     2101 ccccctcata ctccctccca cccacccagc acccgaatca ctcgaagcca gcccaaccac
     2161 acacctgcag gcccgcctgg cccttccagc aaccctgtta gtaacggcaa agaaacccgg
     2221 aggagcagca agagactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     2281 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     2341 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     2401 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2461 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2521 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2581 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     2641 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2701 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     2761 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2821 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     2881 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2941 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     3001 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     3061 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     3121 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     3181 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     3241 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     3301 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     3361 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     3421 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     3481 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     3541 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     3601 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     3661 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     3721 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     3781 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     3841 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3901 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3961 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4021 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4081 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4141 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4201 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4261 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4321 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4381 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4441 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4501 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4561 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4621 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4681 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     4741 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4801 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4861 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4921 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4981 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5041 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5101 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     5161 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     5221 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     5281 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     5341 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     5401 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     5461 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     5521 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     5581 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     5641 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     5701 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     5761 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     5821 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccg
//
