LOCUS       sequence-252204-        4988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human pro-TGF-beta-2 (from L21 to S414) in mammalian 
            cells. With N-t STREP(2x) tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4988)
  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 4988)
  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-STREP
FEATURES             Location/Qualifiers
     source          1..4988
                     /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"
     regulatory      434..443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     440..499
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             join(503..526,527..562,563..586)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     protein_bind    complement(1802..1826)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1827..1846)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1828..1846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1857..1874)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1857..1873)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2278..2412
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2315..2334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2369..2388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2482..2499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2653..3241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2733..2752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3319..3352)
                     /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(3454..4245,4246..4314))
                     /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     4077..4096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        4685..4988
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
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 cggaattccc
      421 cccatcgccc ggggccacca tggagacaga cacactcctg ctatgggtac tgctgctctg
      481 ggttccaggt tccactggtg actggagcca tccacaattc gaaaagggtg gaggttctgg
      541 aggtggaagt ggaggtagcg catggagcca tccacaattc gaaaagctca gcctgtctac
      601 ctgcagcaca ctcgatatgg accagttcat gcgcaagagg atcgaggcga tccgcgggca
      661 gatcctgagc aagctgaagc tcaccagtcc cccagaagac tatcctgagc ccgaggaagt
      721 ccccccggag gtgatttcca tctacaacag caccagggac ttgctccagg agaaggcgag
      781 ccggagggcg gccgcctgcg agcgcgagag gagcgacgaa gagtactacg ccaaggaggt
      841 ttacaaaata gacatgccgc ccttcttccc ctccgaaaat gccatcccgc ccactttcta
      901 cagaccctac ttcagaattg ttcgatttga cgtctcagca atggagaaga atgcttccaa
      961 tttggtgaaa gcagagttca gagtctttcg tttgcagaac ccaaaagcca gagtgcctga
     1021 acaacggatt gagctatatc agattctcaa gtccaaagat ttaacatctc caacccagcg
     1081 ctacatcgac agcaaagttg tgaaaacaag agcagaaggc gaatggctct ccttcgatgt
     1141 aactgatgct gttcatgaat ggcttcacca taaagacagg aacctgggat ttaaaataag
     1201 cttacactgt ccctgctgca cttttgtacc atctaataat tacatcatcc caaataaaag
     1261 tgaagaacta gaagcaagat ttgcaggtat tgatggcacc tccacatata ccagtggtga
     1321 tcagaaaact ataaagtcca ctaggaaaaa aaacagtggg aagaccccac atctcctgct
     1381 aatgttattg ccctcctaca gacttgagtc acaacagacc aaccggcgga agaagcgtgc
     1441 tttggatgcg gcctattgct ttagaaatgt gcaggataat tgctgcctac gtccacttta
     1501 cattgatttc aagagggatc tagggtggaa atggatacac gaacccaaag ggtacaatgc
     1561 caacttctgt gctggagcat gcccgtattt atggagttca gacactcagc acagcagggt
     1621 cctgagctta tataatacca taaatccaga agcatctgct tctccttgct gcgtgtccca
     1681 agatttagaa cctctaacca ttctctacta cattggcaaa acacccaaga ttgaacagct
     1741 ttctaatatg attgtaaagt cttgcaaatg cagctagctc gaggggccca agcttacgcg
     1801 tacccagctt tcttgtacaa agtggtccct atagtgagtc gtattataag ctaggcactg
     1861 gccgtcgttt tacaacgtcg tgactgggaa aactgctagc ttgggatctt tgtgaaggaa
     1921 ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta aagctctaag
     1981 gtaaatataa aatttttaag tgtataatgt gttaaactag ctgcatatgc ttgctgcttg
     2041 agagttttgc ttactgagta tgatttatga aaatattata cacaggagct agtgattcta
     2101 attgtttgtg tattttagat tcacagtccc aaggctcatt tcaggcccct cagtcctcac
     2161 agtctgttca tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2221 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2281 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2341 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2401 catgtctgga tcgatcctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3301 agggattttg gtcatgccat aacttcgtat agcatacatt atacgaagtt atggcatgag
     3361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgccag gggtgggcac
     4381 acatatttga taccagcgat ccctacacag cacataattc aatgcgactt ccctctatcg
     4441 cacatcttag acctttattc tccctccagc acacatcgaa gctgccgagc aagccgttct
     4501 caccagtcca agacctggca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4561 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     4621 tatcatgaca ttaacctata aaaataggcg tattacgagg ccctttcacg cattagatgc
     4681 atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     4741 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     4801 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     4861 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4921 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4981 ttaccatg
//
