LOCUS       sequence-252016-        5026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human pro-TGF-beta-2 (from L21 to S414) in insect cells. 
            With C-t H6x tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5026)
  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 5026)
  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-TGFB2-H6
FEATURES             Location/Qualifiers
     source          1..5026
                     /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             1843..1860
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1903..1920
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1949..1993
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             2015..2038
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      2057..2364
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        2394..2498
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2499..2567,2568..3359)
                     /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(2717..2736)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3530..4118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4019..4038
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4272..4289
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4406..4427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4442..4447,4448..4465,4466..4472)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4480..4496
                     /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     4485..4507
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4504..4520
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4504..4520
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        4541..5023
                     /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 ggccttgtct
      661 acctgcagca cactcgatat ggaccagttc atgcgcaaga ggatcgaggc gatccgcggg
      721 cagatcctga gcaagctgaa gctcaccagt cccccagaag actatcctga gcccgaggaa
      781 gtccccccgg aggtgatttc catctacaac agcaccaggg acttgctcca ggagaaggcg
      841 agccggaggg cggccgcctg cgagcgcgag aggagcgacg aagagtacta cgccaaggag
      901 gtttacaaaa tagacatgcc gcccttcttc ccctccgaaa atgccatccc gcccactttc
      961 tacagaccct acttcagaat tgttcgattt gacgtctcag caatggagaa gaatgcttcc
     1021 aatttggtga aagcagagtt cagagtcttt cgtttgcaga acccaaaagc cagagtgcct
     1081 gaacaacgga ttgagctata tcagattctc aagtccaaag atttaacatc tccaacccag
     1141 cgctacatcg acagcaaagt tgtgaaaaca agagcagaag gcgaatggct ctccttcgat
     1201 gtaactgatg ctgttcatga atggcttcac cataaagaca ggaacctggg atttaaaata
     1261 agcttacact gtccctgctg cacttttgta ccatctaata attacatcat cccaaataaa
     1321 agtgaagaac tagaagcaag atttgcaggt attgatggca cctccacata taccagtggt
     1381 gatcagaaaa ctataaagtc cactaggaaa aaaaacagtg ggaagacccc acatctcctg
     1441 ctaatgttat tgccctccta cagacttgag tcacaacaga ccaaccggcg gaagaagcgt
     1501 gctttggatg cggcctattg ctttagaaat gtgcaggata attgctgcct acgtccactt
     1561 tacattgatt tcaagaggga tctagggtgg aaatggatac acgaacccaa agggtacaat
     1621 gccaacttct gtgctggagc atgcccgtat ttatggagtt cagacactca gcacagcagg
     1681 gtcctgagct tatataatac cataaatcca gaagcatctg cttctccttg ctgcgtgtcc
     1741 caagatttag aacctctaac cattctctac tacattggca aaacacccaa gattgaacag
     1801 ctttctaata tgattgtaaa gtcttgcaaa tgcagcaccg gtcatcatca ccatcaccat
     1861 tgagtttggc aaggccggat ccgttctacc ctgcgtcacg tgcatcacca tcaccatcac
     1921 taataagctt gcggccgcat cttctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     1981 gcacatggac tcgccaccgc cttctggcct cgagcaccac catcaccatc accatcacta
     2041 agtgattaac ctcaggttat acatatattt tgaatttaat taattataca tatattttat
     2101 attatttttg tcttttatta tcgaggggcc gttgttggtg tggggttttg catagaaata
     2161 acaatgggag ttggcgacgt tgctgcgcca acaccacctc ccttccctcc tttcatcatg
     2221 tatctgtaga taaaataaaa tattaaacct aaaaacaaga ccgcgcctat caacaaaatg
     2281 ataggcatta acttgccgct gacgctgtca ctaacgttgg acgatttgcc gactaaacct
     2341 tcatcgccca gtaaccaatc tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2401 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2461 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2521 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2581 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2641 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2701 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2761 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2821 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2881 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2941 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3001 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3061 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3121 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3181 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3241 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3301 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3361 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3421 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3481 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3541 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3601 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3661 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3721 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3781 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3841 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3901 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3961 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4021 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4081 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4141 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4201 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4261 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4321 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4381 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4441 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4501 acacaggaaa cagctatgac catgattacg aattcccggg cgcgtaaaac acaatcaagt
     4561 atgagtcata agctgatgtc atgttttgca cacggctcat aaccgaactg gctttacgag
     4621 tagaattcta cttgtaacgc acgatcagtg gatgatgtca tttgtttttc aaatcgagat
     4681 gatgtcatgt tttgcacacg gctcataaac tcgctttacg agtagaattc tacgtgtaac
     4741 gcacgatcga ttgatgagtc atttgttttg caatatgata tcatacaata tgactcattt
     4801 gtttttcaaa accgaacttg atttacgggt agaattctac ttgtaaagca caatcaaaaa
     4861 gatgatgtca tttgtttttc aaaactgaac tcgctttacg agtagaattc tacgtgtaaa
     4921 acacaatcaa gaaatgatgt catttgttat aaaaataaaa gctgatgtca tgttttgcac
     4981 atggctcata actaaactcg ctttacgggt agaattctac gcgcgt
//
