LOCUS       sequence-257508-        4928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human pro-TGF-beta-2 (from L21 to S414) in mammalian 
            cells. With N-t H8x tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  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 4928)
  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-TGFB2-H8
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..36
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     18..35
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    37..61
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      95..104
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     101..160
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             164..187
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     protein_bind    complement(1403..1427)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1428..1447)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1429..1447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1458..1475)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1458..1474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1467..1489)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    1879..2013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1916..1935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1970..1989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2083..2100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2254..2842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2334..2353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(2920..2953)
                     /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(3055..3846,3847..3915))
                     /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     3678..3697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        4286..4589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4590..4793
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4744..4764
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4790..4814
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     join(4919..4928,1..7)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4919..4928,1..7)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctatgaccat taggcctatt taggtgacac tatagaacaa gtttgtacaa aaaagcaggc
       61 tggtaccggt ccggaattcc ccccatcgcc cggggccacc atggagacag acacactcct
      121 gctatgggta ctgctgctct gggttccagg ttccactggt gaccaccacc atcaccacca
      181 ccatcatctc agcctgtcta cctgcagcac actcgatatg gaccagttca tgcgcaagag
      241 gatcgaggcg atccgcgggc agatcctgag caagctgaag ctcaccagtc ccccagaaga
      301 ctatcctgag cccgaggaag tccccccgga ggtgatttcc atctacaaca gcaccaggga
      361 cttgctccag gagaaggcga gccggagggc ggccgcctgc gagcgcgaga ggagcgacga
      421 agagtactac gccaaggagg tttacaaaat agacatgccg cccttcttcc cctccgaaaa
      481 tgccatcccg cccactttct acagacccta cttcagaatt gttcgatttg acgtctcagc
      541 aatggagaag aatgcttcca atttggtgaa agcagagttc agagtctttc gtttgcagaa
      601 cccaaaagcc agagtgcctg aacaacggat tgagctatat cagattctca agtccaaaga
      661 tttaacatct ccaacccagc gctacatcga cagcaaagtt gtgaaaacaa gagcagaagg
      721 cgaatggctc tccttcgatg taactgatgc tgttcatgaa tggcttcacc ataaagacag
      781 gaacctggga tttaaaataa gcttacactg tccctgctgc acttttgtac catctaataa
      841 ttacatcatc ccaaataaaa gtgaagaact agaagcaaga tttgcaggta ttgatggcac
      901 ctccacatat accagtggtg atcagaaaac tataaagtcc actaggaaaa aaaacagtgg
      961 gaagacccca catctcctgc taatgttatt gccctcctac agacttgagt cacaacagac
     1021 caaccggcgg aagaagcgtg ctttggatgc ggcctattgc tttagaaatg tgcaggataa
     1081 ttgctgccta cgtccacttt acattgattt caagagggat ctagggtgga aatggataca
     1141 cgaacccaaa gggtacaatg ccaacttctg tgctggagca tgcccgtatt tatggagttc
     1201 agacactcag cacagcaggg tcctgagctt atataatacc ataaatccag aagcatctgc
     1261 ttctccttgc tgcgtgtccc aagatttaga acctctaacc attctctact acattggcaa
     1321 aacacccaag attgaacagc tttctaatat gattgtaaag tcttgcaaat gcagctagct
     1381 cgaggggccc aagcttacgc gtacccagct ttcttgtaca aagtggtccc tatagtgagt
     1441 cgtattataa gctaggcact ggccgtcgtt ttacaacgtc gtgactggga aaactgctag
     1501 cttgggatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     1561 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     1621 gctgcatatg cttgctgctt gagagttttg cttactgagt atgatttatg aaaatattat
     1681 acacaggagc tagtgattct aattgtttgt gtattttaga ttcacagtcc caaggctcat
     1741 ttcaggcccc tcagtcctca cagtctgttc atgatcataa tcagccatac cacatttgta
     1801 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1861 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1921 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1981 aaactcatca atgtatctta tcatgtctgg atcgatcctg cattaatgaa tcggccaacg
     2041 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2101 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2161 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2221 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2281 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2341 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2401 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2461 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2521 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2581 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2641 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2701 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2761 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2821 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2881 gtggaacgaa aactcacgtt aagggatttt ggtcatgcca taacttcgta tagcatacat
     2941 tatacgaagt tatggcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3001 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3061 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3121 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3181 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3241 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3301 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3361 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3421 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3481 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3541 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3601 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3661 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3721 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3781 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3841 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3901 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3961 tctcatgcca ggggtgggca cacatatttg ataccagcga tccctacaca gcacataatt
     4021 caatgcgact tccctctatc gcacatctta gacctttatt ctccctccag cacacatcga
     4081 agctgccgag caagccgttc tcaccagtcc aagacctggc atgagcggat acatatttga
     4141 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4201 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtattacgag
     4261 gccctttcac gcattagatg catgtcgtta cataacttac ggtaaatggc ccgcctggct
     4321 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4381 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4441 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4501 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4561 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     4621 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4681 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4741 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     4801 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
     4861 gggaccgatc cagcctccgg actctagcct aggccgcggg acggataaca atttcacaca
     4921 ggaaacag
//
