LOCUS       sequence-401771-        6992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human serum albumin design HSA2, in pET29b with C-terminal 6His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6992)
  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 6992)
  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: HSA2_His
FEATURES             Location/Qualifiers
     source          1..6992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(19..66)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     62..80
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(133..150)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1922..1944
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1959..1983
                     /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)."
     primer_bind     complement(1983..2002)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1984..2002)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2090..2109)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2315..2392
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2393..3475
                     /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     complement(2412..2431)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4284..4475
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4469..4491)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4577..4716
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4632..4651
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4731..4748)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4902..5490)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4982..5001)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5601..5620)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5612..6427
                     /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             5612..6427
                     /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     complement(5685..5704)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6519..6974)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6656..6677)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6868..6887
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tatagttcct cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta
       61 tgctagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
      121 agccggatct cagtggtggt ggtggtggtg ctcgagcagc cccagtgctg cctggcttgc
      181 ggcaatgagc ttcttgccct cctccgcaaa gcatgtttct ttgtcgtcgg ctttacagca
      241 cttttctaca aaggcactga aatcatccat aatcgctttc agctgttctt tcgtcgcttt
      301 tggtttatgc ttcacaagtt ccacaaggaa ggtctgcttt ttgatttggc gttcttcctc
      361 gctgagggtg caaatgtctg catgaaaggt aaacgtttcg gcattgaact ctttcgggac
      421 gtatgtttcg tctacttcaa gggcgctaaa gcatgggcga cgattgacga gggattccgt
      481 gcagcacttg gtaacccgat cattgacagg ggttttttca tggagcacac ataactgatt
      541 cagaacaatg gacagataat cttcgataca tggcatacgc ttagcttctg gaagtttaca
      601 gcatttgctg cccaccttcc ctaagttgcg ggcaacttct accagcgtcg gcgttgacac
      661 ttggggcact ttcttagtat agcgaatcaa taaagcgttc tggaatttgt attcccccag
      721 ttgctcaaac agttcgcagt tttgtttaat gagattttgg ggctcctcga taagaggttt
      781 aaactcatca aaaactttcg aataacattc gtgagggtcc gccgcggcgc aacacttttc
      841 cagcgtggtt tcatacgttt tggctaagcg cagcaacagg gtgacactgt aatccgggtg
      901 acggcgagca tactcgtaaa gaaacatacc taaaaataca tctttcgctt ccgcataatt
      961 cttgcagaca tctttgcttt caatgaaatc cgctgccaga ctaggcagat ccgccggcat
     1021 ttcatcattt tcaacctccg cgatgcaatg agatttctcc agcagaggtt tctcacaaca
     1081 ttctttcagc ttgctagaaa tgctatcctg attttcgcaa atatatttgg ctaaatctgc
     1141 acgatcatcc atgcattcca acaaatcgcc atgacaacat tcggtatgca ctttcgtcaa
     1201 atcagtcacc agtttactca cttccgcaaa ttccgctttc gggaaacgct gtgacagccg
     1261 cgcaatggcc cacgctttaa atgcgcgctc accaaacttc tggagaatgg cacatttgtg
     1321 ccgttgtttg gcggacgagg ctttcccttc ttcgcgaagt tcgtcaagtt tcggaagcaa
     1381 gcaggccgct ttgtctgccg cttgacagca ctctgtaaag gctgccttgt aacgcttagc
     1441 gaagtacagt aattccggag cgtaaaagta tggatgacga cgagcaatct catacaggta
     1501 cttattgaga aacgtttctt cattgtcgtg gaatgcggta cacataacat caggctccgg
     1561 gcgaacgagc cgtgggagat ttggattatc atctttatgc tgcaaaaaac attcattgcg
     1621 ttccggctcc tgtttcgcgc aacaatcggc catttcacca tacgtttcgc ggagtgtagc
     1681 cacagtgcac agtttatcgc cgaaaagcgt gtgcaggctt ttatcgcagt tctctgctga
     1741 ctcatcggcg acacaggttt ttgcaaactc tgttacttcg ttaaccattt tgaccaaatc
     1801 ctcgaacggg cattgttgta aatactgagc gaacgcgatc aacactaacg ctttaaaatt
     1861 ttcttcgccc agatctttaa aacgatgtgc cacctcgctt ttatgtgcat ccatgggtat
     1921 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
     1981 tcccctatag tgagtcgtat taatttcgcg ggatcgagat cgatctcgat cctctacgcc
     2041 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     2101 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     2161 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     2221 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     2281 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     2341 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     2401 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     2461 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     2521 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     2581 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     2641 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     2701 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     2761 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     2821 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     2881 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     2941 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     3001 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     3061 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     3121 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     3181 cgttggtgcg gacatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     3241 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     3301 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     3361 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3481 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     3541 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     3601 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     3661 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     3721 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     3781 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     3841 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     3901 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     3961 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     4021 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     4081 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     4141 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     4201 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     4261 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     4321 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     4381 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     4441 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     4501 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     4561 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     4621 gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata
     4681 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     4741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5341 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     5581 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     5641 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     5701 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     5761 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     5821 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     5881 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     5941 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     6001 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     6061 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     6121 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     6181 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     6241 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     6301 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     6361 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     6421 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     6541 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6601 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6661 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6721 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6781 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6841 aaatccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6901 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6961 gctacagggc gcgtcccatt cgccaatccg ga
//
