LOCUS       sequence-252094-        6499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human N-alpha-2M (from S24 to E908) in insect cells. With 
            C-t H6x tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6499)
  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 6499)
  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: pIE-N-h-alpha-2M-H6
FEATURES             Location/Qualifiers
     source          1..6499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..592
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     CDS             3316..3333
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3376..3393
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3422..3466
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             3488..3511
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      3530..3837
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        3867..3971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3972..4040,4041..4832)
                     /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(4190..4209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5003..5591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5492..5511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5745..5762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5879..5900
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5915..5920,5921..5938,5939..5945)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5953..5969
                     /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     5958..5980
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5977..5993
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5977..5993
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6014..6496
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
ORIGIN
        1 cgatgtcttt gtgatgcgcg cgacattttt gtaggttatt gataaaatga acggatacgt
       61 tgcccgacat tatcattaaa tccttggcgt agaatttgtc gggtccattg tccgtgtgcg
      121 ctagcatgcc cgtaacggac ctcgtacttt tggcttcaaa ggttttgcgc acagacaaaa
      181 tgtgccacac ttgcagctct gcatgtgtgc gcgttaccac aaatcccaac ggcgcagtgt
      241 acttgttgta tgcaaataaa tctcgataaa ggcgcggcgc gcgaatgcag ctgatcacgt
      301 acgctcctcg tgttccgttc aaggacggtg ttatcgacct cagattaatg tttatcggcc
      361 gactgttttc gtatccgctc accaaacgcg tttttgcatt aacattgtat gtcggcggat
      421 gttctatatc taatttgaat aaataaacga taaccgcgtt ggttttagag ggcataataa
      481 aagaaatatt gttatcgtgt tcgccattag ggcagtataa attgacgttc atgttggata
      541 ttgtttcagt tgcaagttga cactggcggc gacaagatcg tgaacaacca agtgactatg
      601 tacaagctca cagtcttcct gatgttcatc gctttcgtca tcatcgctga ggcctcagtc
      661 tctggaaaac cgcagtatat ggttctggtc ccctccctgc tccacactga gaccactgag
      721 aagggctgtg tccttctgag ctacctgaat gagacagtga ctgtaagtgc ttccttggag
      781 tctgtcaggg gaaacaggag cctcttcact gacctggagg cggagaatga cgtactccac
      841 tgtgtcgcct tcgctgtccc aaagtcttca tccaatgagg aggtaatgtt cctcactgtc
      901 caagtgaaag gaccaaccca agaatttaag aagcggacca cagtgatggt taagaacgag
      961 gacagtctgg tctttgtcca gacagacaaa tcaatctaca aaccagggca gacagtgaaa
     1021 tttcgtgttg tctccatgga tgaaaacttt caccccctga atgagttgat tccactagta
     1081 tacattcagg atcccaaagg aaatcgcatc gcacaatggc agagtttcca gttagagggt
     1141 ggcctcaagc aattttcttt tcccctctca tcagagccct tccagggctc ctacaaggtg
     1201 gtggtacaga agaaatcagg tggaaggaca gagcaccctt tcaccgtgga ggaatttgtt
     1261 cttcccaagt ttgaagtaca agtaacagtg ccaaagataa tcaccatctt ggaagaagag
     1321 atgaatgtat cagtgtgtgg cctatacaca tatgggaagc ctgtccctgg acatgtgact
     1381 gtgagcattt gcagaaagta tagtgacgct tccgactgcc acggtgaaga ttcacaggct
     1441 ttctgtgaga aattcagtgg acagctaaac agccatggct gcttctatca gcaagtaaaa
     1501 accaaggtct tccagctgaa gaggaaggag tatgaaatga aacttcacac tgaggcccag
     1561 atccaagaag aaggaacagt ggtggaattg actggaaggc agtccagtga aatcacaaga
     1621 accataacca aactctcatt tgtgaaagtg gactcacact ttcgacaggg aattcccttc
     1681 tttgggcagg tgcgcctagt agatgggaaa ggcgtcccta taccaaataa agtcatattc
     1741 atcagaggaa atgaagcaaa ctattactcc aatgctacca cggatgagca tggccttgta
     1801 cagttctcta tcaacaccac caatgttatg ggtacctctc ttactgttag ggtcaattac
     1861 aaggatcgta gtccctgtta cggctaccag tgggtgtcag aagaacacga agaggcacat
     1921 cacactgctt atcttgtgtt ctccccaagc aagagctttg tccaccttga gcccatgtct
     1981 catgaactac cctgtggcca tactcagaca gtccaggcac attatattct gaatggaggc
     2041 accctgctgg ggctgaagaa gctctccttc tattatctga taatggcaaa gggaggcatt
     2101 gtccgaactg ggactcatgg actgcttgtg aagcaggaag acatgaaggg ccatttttcc
     2161 atctcaatcc ctgtgaagtc agacattgct cctgtcgctc ggttgctcat ctatgctgtt
     2221 ttacctaccg gggacgtgat tggggattct gcaaaatatg atgttgaaaa ttgtctggcc
     2281 aacaaggtgg atttgagctt cagcccatca caaagtctcc cagcctcaca cgcccacctg
     2341 cgagtcacag cggctcctca gtccgtctgc gccctccgtg ctgtggacca aagcgtgctg
     2401 ctcatgaagc ctgatgctga gctctcggcg tcctcggttt acaacctgct accagaaaag
     2461 gacctcactg gcttccctgg gcctttgaat gaccaggacg atgaagactg catcaatcgt
     2521 cataatgtct atattaatgg aatcacatat actccagtat caagtacaaa tgaaaaggat
     2581 atgtacagct tcctagagga catgggctta aaggcattca ccaactcaaa gattcgtaaa
     2641 cccaaaatgt gtccacagct tcaacagtat gaaatgcatg gacctgaagg tctacgtgta
     2701 ggtttttatg agtcagatgt aatgggaaga ggccatgcac gcctggtgca tgttgaagag
     2761 cctcacacgg agaccgtacg aaagtacttc cctgagacat ggatctggga tttggtggtg
     2821 gtaaactcag caggtgtggc tgaggtagga gtaacagtcc ctgacaccat caccgagtgg
     2881 aaggcagggg ccttctgcct gtctgaagat gctggacttg gtatctcttc cactgcctct
     2941 ctccgagcct tccagccctt ctttgtggag ctcacaatgc cttactctgt gattcgtgga
     3001 gaggccttca cactcaaggc cacggtccta aactaccttc ccaaatgcat ccgggtcagt
     3061 gtgcagctgg aagcctctcc cgccttccta gctgtcccag tggagaagga acaagcgcct
     3121 cactgcatct gtgcaaacgg gcggcaaact gtgtcctggg cagtaacccc aaagtcatta
     3181 ggaaatgtga atttcactgt gagcgcagag gcactagagt ctcaagagct gtgtgggact
     3241 gaggtgcctt cagttcctga acacggaagg aaagacacag tcatcaagcc tctgttggtt
     3301 gaacctgaaa ccggtcatca tcaccatcac cattgagttt ggcaaggccg gatccgttct
     3361 accctgcgtc acgtgcatca ccatcaccat cactaataag cttgcggccg catcttctgg
     3421 taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgccac cgccttctgg
     3481 cctcgagcac caccatcacc atcaccatca ctaagtgatt aacctcaggt tatacatata
     3541 ttttgaattt aattaattat acatatattt tatattattt ttgtctttta ttatcgaggg
     3601 gccgttgttg gtgtggggtt ttgcatagaa ataacaatgg gagttggcga cgttgctgcg
     3661 ccaacaccac ctcccttccc tcctttcatc atgtatctgt agataaaata aaatattaaa
     3721 cctaaaaaca agaccgcgcc tatcaacaaa atgataggca ttaacttgcc gctgacgctg
     3781 tcactaacgt tggacgattt gccgactaaa ccttcatcgc ccagtaacca atctagacgt
     3841 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3961 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4021 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4081 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4141 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4201 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4261 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4321 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4381 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4441 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4501 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4561 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4621 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4681 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4741 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4801 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4861 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4921 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4981 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5041 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5101 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5161 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5221 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5281 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5341 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5401 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5461 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5521 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5581 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5641 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5701 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5761 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5821 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5881 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5941 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     6001 acgaattccc gggcgcgtaa aacacaatca agtatgagtc ataagctgat gtcatgtttt
     6061 gcacacggct cataaccgaa ctggctttac gagtagaatt ctacttgtaa cgcacgatca
     6121 gtggatgatg tcatttgttt ttcaaatcga gatgatgtca tgttttgcac acggctcata
     6181 aactcgcttt acgagtagaa ttctacgtgt aacgcacgat cgattgatga gtcatttgtt
     6241 ttgcaatatg atatcataca atatgactca tttgtttttc aaaaccgaac ttgatttacg
     6301 ggtagaattc tacttgtaaa gcacaatcaa aaagatgatg tcatttgttt ttcaaaactg
     6361 aactcgcttt acgagtagaa ttctacgtgt aaaacacaat caagaaatga tgtcatttgt
     6421 tataaaaata aaagctgatg tcatgttttg cacatggctc ataactaaac tcgctttacg
     6481 ggtagaattc tacgcgcgt
//
