LOCUS       sequence-489489-        4726 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  Constitutive expression of human TGF-beta1 (with the human IgK light
            chain secretion sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4726)
  AUTHORS   Edelstein HI, Cosio A, Ezekiel ML, Corcoran WK, Morris AH, Leonard 
            JN
  TITLE     Conversion of natural cytokine receptors into orthogonal synthetic 
            biosensors.
  JOURNAL   Nat Chem Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1.
  PUBMED    40846994
REFERENCE   2  (bases 1 to 4726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2025 Aug 22:10.1038/s41589-025-01986-1. doi: 
            10.1038/s41589-025-01986-1."
COMMENT     Sequence Label: L4278 hIgKLSS_hTGFb1 in pcDNA backbone
FEATURES             Location/Qualifiers
     source          1..4726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1356..1373)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1362..1586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1632..2060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1719..1738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1929..1950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2083..2279
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2130..2265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2192..2211
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2399..2520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2436..2455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2490..2509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2569..2585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2582..2604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2593..2609
                     /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)."
     promoter        complement(join(2617..2623,2624..2641,2642..2647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2662..2683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2800..2817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3051..3070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3730..4521,4522..4590))
                     /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     4353..4372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4591..4695)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagtcccaa gctggctagc
      901 gccaccatgg acatgagagt gcccgcccag ctgctgggcc tgctgctgct gtggctgaga
      961 ggcgccagat gcgccctgga caccaactat tgcttcagct ccacggagaa gaactgctgc
     1021 gtgcggcagc tgtacattga cttccgcaag gacctcggct ggaagtggat ccacgagccc
     1081 aagggctacc atgccaactt ctgcctcggg ccctgcccct acatttggag cctggacacg
     1141 cagtacagca aggtcctggc cctgtacaac cagcataacc cgggcgcctc ggcggcgccg
     1201 tgctgcgtgc cgcaggcgct ggagccgctg cccatcgtgt actacgtggg ccgcaagccc
     1261 aaggtggagc agctgtccaa catgatcgtg cgctcctgca agtgcagcta ggcggccgct
     1321 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     1381 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1441 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1501 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca
     1561 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1621 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1681 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1741 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1801 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1861 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1921 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1981 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2041 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttaggctc aattagtcag
     2101 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2161 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2221 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2281 agctcccggg agcttgtata tccattttcg gaggttgggc ttcggaatcg ttttccggga
     2341 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     2401 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2461 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2521 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     2581 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2641 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2761 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3661 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3841 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     3901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4321 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4681 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
