LOCUS       sequence-403705-        6986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bovine serum albumin design BSA3, in pET29b with C-terminal 
            6His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  AUTHORS   Khersonsky O, Goldsmith M, Zaretsky I, Hamer-Rogotner S, Dym O, 
            Unger T, Yona M, Fridmann-Sirkis Y, Fleishman SJ
  TITLE     Stable Mammalian Serum Albumins Designed for Bacterial Expression.
  JOURNAL   J Mol Biol. 2023 Sep 1;435(17):168191. doi: 
            10.1016/j.jmb.2023.168191. Epub 2023 Jun 28.
  PUBMED    37385581
REFERENCE   2  (bases 1 to 6986)
  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"; journalName: "J Mol 
            Biol."; date: "2023-09-1"; volume: "435(17):168191. doi"; pages: " 
            10.1016/j.jmb.2023.168191. Epub 2023 Jun 28"
COMMENT     Sequence Label: BSA3_His
FEATURES             Location/Qualifiers
     source          1..6986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      42..497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(129..148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     339..360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(589..1404)
                     /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(589..1404)
                     /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     1312..1331
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1396..1415
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1526..2114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2015..2034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2268..2285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2300..2439
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2365..2384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2525..2547
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2541..2732)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3541..4623)
                     /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     4585..4604
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4624..4701)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4907..4926
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5014..5033
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5014..5032
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5033..5057
                     /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             5072..5094
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6860..6877
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6930..6948)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      join(6944..6986,1..5)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
       61 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      121 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      181 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      241 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      301 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      361 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      421 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      481 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      541 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
      601 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      661 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
      721 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
      781 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
      841 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
      901 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
      961 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     1021 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     1081 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     1141 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1201 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1261 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1321 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     1381 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     1441 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2221 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2281 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2341 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2401 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2461 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2521 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2581 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2641 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2701 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2761 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     2821 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     2881 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     2941 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3001 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3061 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3121 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     3181 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     3241 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     3301 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     3361 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     3421 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3481 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3541 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3601 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     3661 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     3721 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     3781 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     3841 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     3901 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     3961 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     4021 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4081 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     4141 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     4201 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     4261 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     4321 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     4381 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     4441 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     4501 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     4561 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     4621 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     4681 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     4741 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     4801 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     4861 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     4921 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     4981 agaggatcga gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt
     5041 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
     5101 catggatacc cataagagtg aaattgccca tcgtttcaaa gaccttggag aagaacactt
     5161 taaagctctc gtgctcattg ccttcgcgca atatctccaa aagtgtcctt ttgatgagtt
     5221 ggtaaagatg gtaaacgatt tgacagagtt tgcgaagaaa tgtgttgcgg atgaatcgca
     5281 tccaggttgc agtaagagcc tgcacactct gtttggagat gaactttgta aagtagagga
     5341 actgcgagaa acctacggag atatggctga ctgttgcagc aaacaggagc ctgaacggaa
     5401 tgaatgcttc ctctcccata aggacgacaa tcccgacttg ccgaaattga agcccgatcc
     5461 gaacactctg tgcaaagaat ttaaagaaga tgaaaagaaa ttttggggca aatacattta
     5521 tgaaatagcc cgccggcatc cgtatttcta tgccccggaa ttactttatt atgcgaagaa
     5581 atataacaaa atctttcaag aatgctgcca ggctgaagat aaagcagctt gcttgctccc
     5641 gaaaattgaa gcgattcggg aaaaagtgaa aacttcttca gctcgccagc gtctgcgttg
     5701 tgcttcatta cagaaatttg gcgaacgggc tctgaaagcc tggtttgtcg cacgtctgtc
     5761 acagaagttt cccaaggctc cgtttgagga aattacgaag ttggtcacag acttaacaaa
     5821 aatttataaa gaatgttgtc acggtgatct gctggaatgt atggatgatc gtgccaagct
     5881 ggtaaagtac atttgtgata atcaagatac gataagttca aagttaaagg agtgttgcga
     5941 taaaccactt ttagagagaa gtcactgtat tgcggaagtc gaggatgatg atatcccgga
     6001 ggacctccct cctcttgtag cggattttct ggaggataaa gatgtctgta aaaattatca
     6061 ggaagataaa gacgcttttc ttggccgttt tttatacgaa tatagcagac gtcaccccga
     6121 atatgctgtg tctttgttgc ttcgccttgc aaaggagtac gaagcaactc tggaggaatg
     6181 ctgtaaaaaa gacgatccac acgcttgtta tagtcgggtc tttgagaaac tgaagaaaca
     6241 tatagatgag ccgcaagaac tgataaaaca gaactgcgac cagtttgaga agctggggga
     6301 atacggcttc caaaatgcat tgataattag atacactaaa aagatgccgc aagtgccgac
     6361 ggataccctc gttgagataa ctcggagttt aggaaaagta ggcacacggt gttgccagct
     6421 tccggaagaa aaacgcttgc cttgcactga ggactattta tccctcattt taaaccggct
     6481 gtgcgtactt catgaaaaga ctccagtgag tgagaaggtt accaaatgtt gcactgaaag
     6541 cctggtgaat agacgtcact gtttttcagc gttaactcca gatgaaactt atgtccccaa
     6601 accttttgat cctaaattgt tcacctttca cgctgatata tgtaccttac ctcccgaaga
     6661 aaaacagatc aagaagcaga aatttttggt ggaacttttg aaacataagc cgaaagctac
     6721 cgaagaacag ttgaaaacta ttatggaaaa ctttacagct atggtcgata agtgttgtaa
     6781 agctgaggat aaagaagctt gttttgccga agagggacca aaactgattg agaaaaccca
     6841 agcggccctg gctctcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
     6901 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     6961 ggcctctaaa cgggtcttga ggggtt
//
