LOCUS       sequence-252203-        8658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human alpha-2M (from G27 to S1468) in mammalian cells. 
            With C-t H6x tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8658)
  AUTHORS   Marino-Puertas L, Del Amo-Maestro L, Taules M, Gomis-Ruth FX, Goulas
            T
  TITLE     Recombinant production of human alpha2-macroglobulin variants and 
            interaction studies with recombinant G-related alpha2-macroglobulin 
            binding protein and latent transforming growth factor-beta2.
  JOURNAL   Sci Rep. 2019 Jun 24;9(1):9186. doi: 10.1038/s41598-019-45712-z.
  PUBMED    31235767
REFERENCE   2  (bases 1 to 8658)
  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: "Sci Rep."; 
            date: "2019-06-24"; volume: "9(1):9186. doi"; pages: " 
            10.1038/s41598-019-45712-z"
COMMENT     Sequence Label: pS6-h-alpha-2M-H6
FEATURES             Location/Qualifiers
     source          1..8658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        357..375
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     357..374
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    376..400
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     426..485
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4815..4832
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(4880..4904)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4905..4924)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4906..4924)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4935..4952)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4935..4951)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4944..4966)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5356..5490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5393..5412)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5447..5466
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5560..5577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5731..6319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5811..5830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(6397..6430)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(6532..7323,7324..7392))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7155..7174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7679..8133)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7816..7837)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8027..8046
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(8263..8367)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcc
      301 taggccgcgg gacggataac aatttcacac aggaaacagc tatgaccatt aggcctattt
      361 aggtgacact atagaacaag tttgtacaaa aaagcaggct ggtaccggtc cggaatcccc
      421 ccgggatgga gacagacaca ctcctgctat gggtactgct gctctgggtt ccaggttcca
      481 ctggtgacgg aaaaccgcag tatatggttc tggtcccctc cctgctccac actgagacca
      541 ctgagaaggg ctgtgtcctt ctgagctacc tgaatgagac agtgactgta agtgcttcct
      601 tggagtctgt caggggaaac aggagcctct tcactgacct ggaggcggag aatgacgtac
      661 tccactgtgt cgccttcgct gtcccaaagt cttcatccaa tgaggaggta atgttcctca
      721 ctgtccaagt gaaaggacca acccaagaat ttaagaagcg gaccacagtg atggttaaga
      781 acgaggacag tctggtcttt gtccagacag acaaatcaat ctacaaacca gggcagacag
      841 tgaaatttcg tgttgtctcc atggatgaaa actttcaccc cctgaatgag ttgattccac
      901 tagtatacat tcaggatccc aaaggaaatc gcatcgcaca atggcagagt ttccagttag
      961 agggtggcct caagcaattt tcttttcccc tctcatcaga gcccttccag ggctcctaca
     1021 aggtggtggt acagaagaaa tcaggtggaa ggacagagca ccctttcacc gtggaggaat
     1081 ttgttcttcc caagtttgaa gtacaagtaa cagtgccaaa gataatcacc atcttggaag
     1141 aagagatgaa tgtatcagtg tgtggcctat acacatatgg gaagcctgtc cctggacatg
     1201 tgactgtgag catttgcaga aagtatagtg acgcttccga ctgccacggt gaagattcac
     1261 aggctttctg tgagaaattc agtggacagc taaacagcca tggctgcttc tatcagcaag
     1321 taaaaaccaa ggtcttccag ctgaagagga aggagtatga aatgaaactt cacactgagg
     1381 cccagatcca agaagaagga acagtggtgg aattgactgg aaggcagtcc agtgaaatca
     1441 caagaaccat aaccaaactc tcatttgtga aagtggactc acactttcga cagggaattc
     1501 ccttctttgg gcaggtgcgc ctagtagatg ggaaaggcgt ccctatacca aataaagtca
     1561 tattcatcag aggaaatgaa gcaaactatt actccaatgc taccacggat gagcatggcc
     1621 ttgtacagtt ctctatcaac accaccaatg ttatgggtac ctctcttact gttagggtca
     1681 attacaagga tcgtagtccc tgttacggct accagtgggt gtcagaagaa cacgaagagg
     1741 cacatcacac tgcttatctt gtgttctccc caagcaagag ctttgtccac cttgagccca
     1801 tgtctcatga actaccctgt ggccatactc agacagtcca ggcacattat attctgaatg
     1861 gaggcaccct gctggggctg aagaagctct ccttctatta tctgataatg gcaaagggag
     1921 gcattgtccg aactgggact catggactgc ttgtgaagca ggaagacatg aagggccatt
     1981 tttccatctc aatccctgtg aagtcagaca ttgctcctgt cgctcggttg ctcatctatg
     2041 ctgttttacc taccggggac gtgattgggg attctgcaaa atatgatgtt gaaaattgtc
     2101 tggccaacaa ggtggatttg agcttcagcc catcacaaag tctcccagcc tcacacgccc
     2161 acctgcgagt cacagcggct cctcagtccg tctgcgccct ccgtgctgtg gaccaaagcg
     2221 tgctgctcat gaagcctgat gctgagctct cggcgtcctc ggtttacaac ctgctaccag
     2281 aaaaggacct cactggcttc cctgggcctt tgaatgacca ggacgatgaa gactgcatca
     2341 atcgtcataa tgtctatatt aatggaatca catatactcc agtatcaagt acaaatgaaa
     2401 aggatatgta cagcttccta gaggacatgg gcttaaaggc attcaccaac tcaaagattc
     2461 gtaaacccaa aatgtgtcca cagcttcaac agtatgaaat gcatggacct gaaggtctac
     2521 gtgtaggttt ttatgagtca gatgtaatgg gaagaggcca tgcacgcctg gtgcatgttg
     2581 aagagcctca cacggagacc gtacgaaagt acttccctga gacatggatc tgggatttgg
     2641 tggtggtaaa ctcagcaggt gtggctgagg taggagtaac agtccctgac accatcaccg
     2701 agtggaaggc aggggccttc tgcctgtctg aagatgctgg acttggtatc tcttccactg
     2761 cctctctccg agccttccag cccttctttg tggagctcac aatgccttac tctgtgattc
     2821 gtggagaggc cttcacactc aaggccacgg tcctaaacta ccttcccaaa tgcatccggg
     2881 tcagtgtgca gctggaagcc tctcccgcct tcctagctgt cccagtggag aaggaacaag
     2941 cgcctcactg catctgtgca aacgggcggc aaactgtgtc ctgggcagta accccaaagt
     3001 cattaggaaa tgtgaatttc actgtgagcg cagaggcact agagtctcaa gagctgtgtg
     3061 ggactgaggt gccttcagtt cctgaacacg gaaggaaaga cacagtcatc aagcctctgt
     3121 tggttgaacc tgaaggacta gagaaggaaa caacattcaa ctccctactt tgtccatcag
     3181 gtggtgaggt ttctgaagaa ttatccctga aactgccacc aaatgtggta gaagaatctg
     3241 cccgagcttc tgtctcagtt ttgggagaca tattaggctc tgccatgcaa aacacacaaa
     3301 atcttctcca gatgccctat ggctgtggag agcagaatat ggtcctcttt gctcctaaca
     3361 tctatgtact ggattatcta aatgaaacac agcagcttac tccagaggtc aagtccaagg
     3421 ccattggcta tctcaacact ggttaccaga gacagttgaa ctacaaacac tatgatggct
     3481 cctacagcac ctttggggag cgatatggca ggaaccaggg caacacctgg ctcacagcct
     3541 ttgttctgaa gacttttgcc caagctcgag cctacatctt catcgatgaa gcacacatta
     3601 cccaagccct catatggctc tcccagaggc agaaggacaa tggctgtttc aggagctctg
     3661 ggtcactgct caacaatgcc ataaagggag gagtagaaga tgaagtgacc ctctccgcct
     3721 atatcaccat cgcccttctg gagattcctc tcacagtcac tcaccctgtt gtccgcaatg
     3781 ccctgttttg cctggagtca gcctggaaga cagcacaaga aggggaccat ggcagccatg
     3841 tatataccaa agcactgctg gcctatgctt ttgccctggc aggtaaccag gacaagagga
     3901 aggaagtact caagtcactt aatgaggaag ctgtgaagaa agacaactct gtccattggg
     3961 agcgccctca gaaacccaag gcaccagtgg ggcattttta cgaaccccag gctccctctg
     4021 ctgaggtgga gatgacatcc tatgtgctcc tcgcttatct cacggcccag ccagccccaa
     4081 cctcggagga cctgacctct gcaaccaaca tcgtgaagtg gatcacgaag cagcagaatg
     4141 cccagggcgg tttctcctcc acccaggaca cagtggtggc tctccatgct ctgtccaaat
     4201 atggagcagc cacatttacc aggactggga aggctgcaca ggtgactatc cagtcttcag
     4261 ggacattttc cagcaaattc caagtggaca acaacaaccg cctgttactg cagcaggtct
     4321 cattgccaga gctgcctggg gaatacagca tgaaagtgac aggagaagga tgtgtctacc
     4381 tccagacatc cttgaaatac aatattctcc cagaaaagga agagttcccc tttgctttag
     4441 gagtgcagac tctgcctcaa acttgtgatg aacccaaagc ccacaccagc ttccaaatct
     4501 ccctaagtgt cagttacaca gggagccgct ctgcctccaa catggcgatc gttgatgtga
     4561 agatggtctc tggcttcatt cccctgaagc caacagtgaa aatgcttgaa agatctaacc
     4621 atgtgagccg gacagaagtc agcagcaacc atgtcttgat ttaccttgat aaggtgtcaa
     4681 atcagacact gagcttgttc ttcacggttc tgcaagatgt cccagtaaga gatctgaaac
     4741 cagccatagt gaaagtctat gattactacg agacggatga gtttgcaatt gctgagtaca
     4801 atgctccttg cagccaccat caccatcacc attaggcggc cgctctagag tatccctcga
     4861 ggggcccaag cttacgcgta cccagctttc ttgtacaaag tggtccctat agtgagtcgt
     4921 attataagct aggcactggc cgtcgtttta caacgtcgtg actgggaaaa ctgctagctt
     4981 gggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca
     5041 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactagct
     5101 gcatatgctt gctgcttgag agttttgctt actgagtatg atttatgaaa atattataca
     5161 caggagctag tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc
     5221 aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag
     5281 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5341 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5401 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5461 ctcatcaatg tatcttatca tgtctggatc gatcctgcat taatgaatcg gccaacgcgc
     5521 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6361 gaacgaaaac tcacgttaag ggattttggt catgccataa cttcgtatag catacattat
     6421 acgaagttat ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7441 catgccaggg gtgggcacac atatttgata ccagcgatcc ctacacagca cataattcaa
     7501 tgcgacttcc ctctatcgca catcttagac ctttattctc cctccagcac acatcgaagc
     7561 tgccgagcaa gccgttctca ccagtccaag acctggcatg agcggataca tatttgaatg
     7621 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
     7681 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     7741 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     7801 gggttgagtg ttgttccagt ttggaacaag agtccactat taagaacgtg gactccaacg
     7861 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     7921 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7981 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     8041 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     8101 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     8161 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagccaata cgcaaaccgc
     8221 ctctccccgc gcgttggccg attcattaat gcagagcttg caattcgcgc tttttcaata
     8281 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8341 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     8401 agaaaccatt attatcatga cattaaccta taaaaatagg cgtattacga ggccctttca
     8461 cgcattagat gcatgtcgtt acataactta cggtaaatgg cccgcctggc agtacatcaa
     8521 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     8581 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     8641 gtcatcgcta ttaccatg
//
