LOCUS       sequence-252007-        6618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human N-alpha-2M (from G27 to A788) in mammalian cells. 
            With C-t H6x tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6618)
  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 6618)
  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-N-h-alpha-2M-H6
FEATURES             Location/Qualifiers
     source          1..6618
                     /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             5351..5368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(5416..5440)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5441..5460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5442..5460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5471..5488)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5471..5487)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5480..5502)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    5892..6026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5929..5948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5983..6002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6152..6606
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6239..6258)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6448..6469
                     /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 tgacggaaaa ccgcagtata tggttctggt cccctccctg ctccacactg agaccactga
     3121 gaagggctgt gtccttctga gctacctgaa tgagacagtg actgtaagtg cttccttgga
     3181 gtctgtcagg ggaaacagga gcctcttcac tgacctggag gcggagaatg acgtactcca
     3241 ctgtgtcgcc ttcgctgtcc caaagtcttc atccaatgag gaggtaatgt tcctcactgt
     3301 ccaagtgaaa ggaccaaccc aagaatttaa gaagcggacc acagtgatgg ttaagaacga
     3361 ggacagtctg gtctttgtcc agacagacaa atcaatctac aaaccagggc agacagtgaa
     3421 atttcgtgtt gtctccatgg atgaaaactt tcaccccctg aatgagttga ttccactagt
     3481 atacattcag gatcccaaag gaaatcgcat cgcacaatgg cagagtttcc agttagaggg
     3541 tggcctcaag caattttctt ttcccctctc atcagagccc ttccagggct cctacaaggt
     3601 ggtggtacag aagaaatcag gtggaaggac agagcaccct ttcaccgtgg aggaatttgt
     3661 tcttcccaag tttgaagtac aagtaacagt gccaaagata atcaccatct tggaagaaga
     3721 gatgaatgta tcagtgtgtg gcctatacac atatgggaag cctgtccctg gacatgtgac
     3781 tgtgagcatt tgcagaaagt atagtgacgc ttccgactgc cacggtgaag attcacaggc
     3841 tttctgtgag aaattcagtg gacagctaaa cagccatggc tgcttctatc agcaagtaaa
     3901 aaccaaggtc ttccagctga agaggaagga gtatgaaatg aaacttcaca ctgaggccca
     3961 gatccaagaa gaaggaacag tggtggaatt gactggaagg cagtccagtg aaatcacaag
     4021 aaccataacc aaactctcat ttgtgaaagt ggactcacac tttcgacagg gaattccctt
     4081 ctttgggcag gtgcgcctag tagatgggaa aggcgtccct ataccaaata aagtcatatt
     4141 catcagagga aatgaagcaa actattactc caatgctacc acggatgagc atggccttgt
     4201 acagttctct atcaacacca ccaatgttat gggtacctct cttactgtta gggtcaatta
     4261 caaggatcgt agtccctgtt acggctacca gtgggtgtca gaagaacacg aagaggcaca
     4321 tcacactgct tatcttgtgt tctccccaag caagagcttt gtccaccttg agcccatgtc
     4381 tcatgaacta ccctgtggcc atactcagac agtccaggca cattatattc tgaatggagg
     4441 caccctgctg gggctgaaga agctctcctt ctattatctg ataatggcaa agggaggcat
     4501 tgtccgaact gggactcatg gactgcttgt gaagcaggaa gacatgaagg gccatttttc
     4561 catctcaatc cctgtgaagt cagacattgc tcctgtcgct cggttgctca tctatgctgt
     4621 tttacctacc ggggacgtga ttggggattc tgcaaaatat gatgttgaaa attgtctggc
     4681 caacaaggtg gatttgagct tcagcccatc acaaagtctc ccagcctcac acgcccacct
     4741 gcgagtcaca gcggctcctc agtccgtctg cgccctccgt gctgtggacc aaagcgtgct
     4801 gctcatgaag cctgatgctg agctctcggc gtcctcggtt tacaacctgc taccagaaaa
     4861 ggacctcact ggcttccctg ggcctttgaa tgaccaggac gatgaagact gcatcaatcg
     4921 tcataatgtc tatattaatg gaatcacata tactccagta tcaagtacaa atgaaaagga
     4981 tatgtacagc ttcctagagg acatgggctt aaaggcattc accaactcaa agattcgtaa
     5041 acccaaaatg tgtccacagc ttcaacagta tgaaatgcat ggacctgaag gtctacgtgt
     5101 aggtttttat gagtcagatg taatgggaag aggccatgca cgcctggtgc atgttgaaga
     5161 gcctcacacg gagaccgtac gaaagtactt ccctgagaca tggatctggg atttggtggt
     5221 ggtaaactca gcaggtgtgg ctgaggtagg agtaacagtc cctgacacca tcaccgagtg
     5281 gaaggcaggg gccttctgcc tgtctgaaga tgctggactt ggtatctctt ccactgcctc
     5341 tctccgagcc caccatcacc atcaccatta ggcggccgct ctagagtatc cctcgagggg
     5401 cccaagctta cgcgtaccca gctttcttgt acaaagtggt ccctatagtg agtcgtatta
     5461 taagctaggc actggccgtc gttttacaac gtcgtgactg ggaaaactgc tagcttggga
     5521 tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga
     5581 tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctagctgcat
     5641 atgcttgctg cttgagagtt ttgcttactg agtatgattt atgaaaatat tatacacagg
     5701 agctagtgat tctaattgtt tgtgtatttt agattcacag tcccaaggct catttcaggc
     5761 ccctcagtcc tcacagtctg ttcatgatca taatcagcca taccacattt gtagaggttt
     5821 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     5881 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5941 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     6001 tcaatgtatc ttatcatgtc tggatcgatc ctgcattaat gaatcggcca acgcgcgggg
     6061 agaggcggtt tgcgtattgg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     6121 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     6181 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6241 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6301 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6361 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6421 ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga acaacactca
     6481 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6541 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6601 caatttaggt ggcacttt
//
