LOCUS       sequence-487919-        7129 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Constitutive expression of intC-ZF9 split synTF component.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7129)
  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 7129)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 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: L4873 intC-ZF9 in PolyTX-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..7129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        359..1541
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          590..1532
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1489..1509
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1543..1567
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2110..2130
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2143..2169
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2206..2430
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             2573..3367
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2627..2646)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3237..3256
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3398..3492
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     enhancer        3513..3892
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3893..4096
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4047..4067
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4141..4160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4141..4159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4179..4188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4932..5156
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5202..5630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5289..5308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5499..5520
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5653..5849
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5700..5835
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5762..5781
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5969..6090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6006..6025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6060..6079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6139..6155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6139..6155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6152..6174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6163..6179
                     /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(6187..6193,6194..6211,6212..6217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6232..6253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6370..6387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6541..7129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6621..6640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag cgatcgaata
      181 ttagatctga gacccaattg acgcgtgagt aattcataca aaaggactcg cccctgcctt
      241 ggggaatccc agggaccgtc gttaaactcc cactaacgta gaacccagag atcgctgcgt
      301 tcccgccccc tcacccgccc gctctcgtca tcactgaggt ggagaagagc atgcgtgagg
      361 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
      421 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
      481 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
      541 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
      601 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
      661 ttccacgccc ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      721 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      781 ggcttgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      841 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      901 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      961 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     1021 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1081 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1141 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1201 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1261 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1321 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1381 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1441 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1501 cagtggttca aagttttttt cttccatttc aggtgtcgtg agagctcgtt tagtgaaccg
     1561 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagag aacgggaccg
     1621 atccaaattc tcgaggctag cgccaccatg atgctgaaga agatcctgaa gatcgaggag
     1681 ctggacgaga gagagctgat cgacatcgag gtgagcggca accacctgtt ctacgccaac
     1741 gacatcctga cccacaacag cggatccggt ggtggcgggt ctggaggagg aggtagtggt
     1801 ggaggtggct ctggtaccgg tggagctaga cccggggagc gccccttcca gtgtcgcatt
     1861 tgcatgcgga acttttcgac cggccagatc cttgaccgcc atacccgtac tcataccggt
     1921 gaaaaaccgt ttcagtgtcg gatctgtatg cgaaatttct ccgtggcgca cagcttgaag
     1981 aggcatctac gtacgcacac cggcgagaag ccattccaat gccgaatatg catgcgcaac
     2041 ttcagtgacc ccagcaacct gcggcgccac ctaaaaaccc acctgagagg atctgaattc
     2101 ggctctggcc ctaagaagaa gcgcaaagtt ggctctggca tgtacccata cgacgtacca
     2161 gattacgctt aggcggccgc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     2221 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2281 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2341 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gtcaatagca
     2401 ggcatgctgg ggatgcggtg ggctctatgg gacgtcggaa ttgcaagttg ggccgccctc
     2461 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     2521 atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     2581 acaagatgga ttgcacgcag gttctccggc ggcttgggtg gagaggctat tcggctatga
     2641 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     2701 tcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     2761 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcggctg tgctcgacgt
     2821 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     2881 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     2941 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3001 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3061 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgtatgcccg acggcgagga
     3121 tctcgtcgtg acccacggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3181 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3241 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3301 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3361 cttctgagcg ggactctggg gttcgagaat tggcttggac tcctgttgat agatccagta
     3421 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     3481 attggtgaga atccaagcac tagccatgac cagacattga ttattgacta gttattaata
     3541 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3601 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3661 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3721 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3781 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3841 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3901 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3961 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4021 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4081 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4141 taatacgact cactataggg agtcccaagc tgactagcgc caccatggtt tccaagggcg
     4201 aagaactgat caaagaaaac atgcacatga agctctacat ggaaggcacc gtggacaacc
     4261 accacttcaa gtgtaccagc gaaggcgagg gcaagcctta cgagggcacc cagaccatga
     4321 gaatcaaggt ggtggaaggc ggccctctgc ctttcgcctt tgatatcctg gccaccagct
     4381 ttctgtacgg cagcaagacc ttcatcaatc acacccaggg catccccgat ttcttcaagc
     4441 agagcttccc cgagggcttc acctgggaga gagtgaccac atacgaggat ggcggcgtgc
     4501 tgacagccac acaggataca agcctccagg acggctgcct gatctacaac gtgaagatcc
     4561 ggggcgtgaa cttcaccagc aacggccccg tgatgcagaa gaaaaccctt ggctgggaag
     4621 ccttcaccga gacactgtat cctgccgatg gcggcctgga aggcagaaac gatatggccc
     4681 tgaagctcgt cggcggctct cacctgattg ccaatgccaa gaccacctac agaagcaaga
     4741 agcccgccaa gaacctgaag atgcccggcg tgtactacgt ggactaccgg ctggaaagaa
     4801 tcaaagaggc caacaacgag acatacgtgg aacagcacga ggtggccgtg gccagatact
     4861 gtgatctgcc ttctaagctg ggccacaagc tgaactgagc ggccaagttt aaacccgctg
     4921 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4981 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     5041 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     5101 gggggaggat tgggaagtca atagcaggca tgctggggat gcggtgggct ctatggcttc
     5161 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5221 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5281 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5341 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5401 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5461 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5521 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5581 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5641 aatgtgtgtc agttaggctc aattagtcag caaccatagt cccgccccta actccgccca
     5701 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5761 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5821 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5881 gaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5941 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6001 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6061 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6121 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6181 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6241 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6301 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6361 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6421 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6481 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6541 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6601 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6661 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6721 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6781 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6841 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6901 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6961 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7021 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7081 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa
//
