LOCUS       sequence-252013-        6372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human C-alpha-2M (from F789 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 6372)
  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 6372)
  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-C-h-alpha-2M-H6
FEATURES             Location/Qualifiers
     source          1..6372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(275..343,344..1135)
                     /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     complement(493..512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1237..1270
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      1348..1936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1837..1856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2090..2107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(2181..2285)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2577..2780
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2731..2751
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2777..2801
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2906..2922
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2906..2922
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2933..2951
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2933..2950
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    2952..2976
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     3002..3061
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5105..5122
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(5170..5194)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5195..5214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5196..5214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5225..5242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5225..5241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5234..5256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5646..5780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5683..5702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5737..5756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5906..6360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5993..6012)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6202..6223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
       61 tccgctcatg ccaggtcttg gactggtgag aacggcttgc tcggcagctt cgatgtgtgc
      121 tggagggaga ataaaggtct aagatgtgcg atagagggaa gtcgcattga attatgtgct
      181 gtgtagggat cgctggtatc aaatatgtgt gcccacccct ggcatgagac aataaccctg
      241 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      301 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      361 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      421 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      481 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
      541 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
      601 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
      661 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
      721 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
      781 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
      841 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
      901 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
      961 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1021 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1081 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1141 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1201 gatctaggtg aagatccttt ttgataatct catgccataa cttcgtataa tgtatgctat
     1261 acgaagttat ggcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2101 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2161 tcattaatgc agagcttgca attcgcgctt tttcaatatt attgaagcat ttatcagggt
     2221 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2281 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     2341 ttaacctata aaaataggcg tattacgagg ccctttcacg cattagatgc atgtcgttac
     2401 ataacttacg gtaaatggcc cgcctggcag tacatcaagt gtatcatatg ccaagtacgc
     2461 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2521 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     2581 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     2641 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     2701 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     2761 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     2821 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagcctagg
     2881 ccgcgggacg gataacaatt tcacacagga aacagctatg accattaggc ctatttaggt
     2941 gacactatag aacaagtttg tacaaaaaag caggctggta ccggtccgga atccccccgg
     3001 gatggagaca gacacactcc tgctatgggt actgctgctc tgggttccag gttccactgg
     3061 tgacttccag cccttctttg tggagctcac aatgccttac tctgtgattc gtggagaggc
     3121 cttcacactc aaggccacgg tcctaaacta ccttcccaaa tgcatccggg tcagtgtgca
     3181 gctggaagcc tctcccgcct tcctagctgt cccagtggag aaggaacaag cgcctcactg
     3241 catctgtgca aacgggcggc aaactgtgtc ctgggcagta accccaaagt cattaggaaa
     3301 tgtgaatttc actgtgagcg cagaggcact agagtctcaa gagctgtgtg ggactgaggt
     3361 gccttcagtt cctgaacacg gaaggaaaga cacagtcatc aagcctctgt tggttgaacc
     3421 tgaaggacta gagaaggaaa caacattcaa ctccctactt tgtccatcag gtggtgaggt
     3481 ttctgaagaa ttatccctga aactgccacc aaatgtggta gaagaatctg cccgagcttc
     3541 tgtctcagtt ttgggagaca tattaggctc tgccatgcaa aacacacaaa atcttctcca
     3601 gatgccctat ggctgtggag agcagaatat ggtcctcttt gctcctaaca tctatgtact
     3661 ggattatcta aatgaaacac agcagcttac tccagaggtc aagtccaagg ccattggcta
     3721 tctcaacact ggttaccaga gacagttgaa ctacaaacac tatgatggct cctacagcac
     3781 ctttggggag cgatatggca ggaaccaggg caacacctgg ctcacagcct ttgttctgaa
     3841 gacttttgcc caagctcgag cctacatctt catcgatgaa gcacacatta cccaagccct
     3901 catatggctc tcccagaggc agaaggacaa tggctgtttc aggagctctg ggtcactgct
     3961 caacaatgcc ataaagggag gagtagaaga tgaagtgacc ctctccgcct atatcaccat
     4021 cgcccttctg gagattcctc tcacagtcac tcaccctgtt gtccgcaatg ccctgttttg
     4081 cctggagtca gcctggaaga cagcacaaga aggggaccat ggcagccatg tatataccaa
     4141 agcactgctg gcctatgctt ttgccctggc aggtaaccag gacaagagga aggaagtact
     4201 caagtcactt aatgaggaag ctgtgaagaa agacaactct gtccattggg agcgccctca
     4261 gaaacccaag gcaccagtgg ggcattttta cgaaccccag gctccctctg ctgaggtgga
     4321 gatgacatcc tatgtgctcc tcgcttatct cacggcccag ccagccccaa cctcggagga
     4381 cctgacctct gcaaccaaca tcgtgaagtg gatcacgaag cagcagaatg cccagggcgg
     4441 tttctcctcc acccaggaca cagtggtggc tctccatgct ctgtccaaat atggagcagc
     4501 cacatttacc aggactggga aggctgcaca ggtgactatc cagtcttcag ggacattttc
     4561 cagcaaattc caagtggaca acaacaaccg cctgttactg cagcaggtct cattgccaga
     4621 gctgcctggg gaatacagca tgaaagtgac aggagaagga tgtgtctacc tccagacatc
     4681 cttgaaatac aatattctcc cagaaaagga agagttcccc tttgctttag gagtgcagac
     4741 tctgcctcaa acttgtgatg aacccaaagc ccacaccagc ttccaaatct ccctaagtgt
     4801 cagttacaca gggagccgct ctgcctccaa catggcgatc gttgatgtga agatggtctc
     4861 tggcttcatt cccctgaagc caacagtgaa aatgcttgaa agatctaacc atgtgagccg
     4921 gacagaagtc agcagcaacc atgtcttgat ttaccttgat aaggtgtcaa atcagacact
     4981 gagcttgttc ttcacggttc tgcaagatgt cccagtaaga gatctgaaac cagccatagt
     5041 gaaagtctat gattactacg agacggatga gtttgcaatt gctgagtaca atgctccttg
     5101 cagccaccat caccatcacc attaggcggc cgctctagag tatccctcga ggggcccaag
     5161 cttacgcgta cccagctttc ttgtacaaag tggtccctat agtgagtcgt attataagct
     5221 aggcactggc cgtcgtttta caacgtcgtg actgggaaaa ctgctagctt gggatctttg
     5281 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     5341 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactagct gcatatgctt
     5401 gctgcttgag agttttgctt actgagtatg atttatgaaa atattataca caggagctag
     5461 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
     5521 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
     5581 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     5641 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5701 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5761 tatcttatca tgtctggatc gatcctgcat taatgaatcg gccaacgcgc ggggagaggc
     5821 ggtttgcgta ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     5881 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5941 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6001 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6061 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6121 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6181 ttggagtcca cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6241 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6301 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6361 aggtggcact tt
//
