LOCUS       sequence-467420-        8143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac donor plasmid, secreted CCL20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8143)
  AUTHORS   Zhang X, Mille-Fragoso LS, Kaseniit KE, Lee AP, Zhang M, Call CC, Hu
            Y, Xie Y, Gao XJ
  TITLE     Post-transcriptional modular synthetic receptors.
  JOURNAL   Nat Chem Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w.
  PUBMED    40155716
REFERENCE   2  (bases 1 to 8143)
  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: "Nat Chem 
            Biol. 2025 Mar 28. doi: 10.1038/s41589-025-01872-w."
COMMENT     Sequence Label: XZ091 (PB) CMV-sCCL20 (PuroR-BFP)
FEATURES             Location/Qualifiers
     source          1..8143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1744..1766
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1758..1775
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1759..1775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1905..1967
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2458..2837
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2838..3041
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2992..3012
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3043..3061
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3064..3082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3092..3116
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3535..3564
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3620..3637)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3626..3833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3866..3886)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3871..4200
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4051..4186
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4113..4132
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4249..4845
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4249..4268)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4745..4765
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4846..4902
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4909..5619
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(5682..5702)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5744..5865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5781..5800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5835..5854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6628..6662
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7153..7169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7153..7169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7166..7188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7177..7193
                     /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(7201..7207,7208..7225,7226..7231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7246..7267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7384..7401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7555..8143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7635..7654)
                     /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 ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1741 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt
     1801 ttgaacccgt ggaggacggg cagactcgcg gtgcaaatgt gttttacagc gtgatggagc
     1861 agatgaagat gctcgacacg ctgcagaaca cgcagctaga ttaaccctag aaagataatc
     1921 atattgtgac gtacgttaaa gataatcatg tgtaaaattg acgcatgtgt tttatcggtc
     1981 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
     2041 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
     2101 caaaaactca aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc
     2161 aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg
     2221 gggctccgct ccggtccggc gctccccccg catcccagag ccggcagcgt gcggggacag
     2281 cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt
     2341 gagcctgcag acacctgggg ggatacgggg aaaaggcctc cacggccaca attgcatgaa
     2401 gaatctgctt agggttaggc gttttgcgct gcttcgtacg ggccagatat acgcgttgac
     2461 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2521 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2581 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2641 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2701 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2761 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2821 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2881 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2941 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3001 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag
     3061 atctccctat cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct
     3121 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     3181 ggactctagc gtttaaactt aagcttcgcc accatgtgct gtaccaagag tttgctcctg
     3241 gctgctttga tgtcagtgct gctactccac ctctgcggcg aatcagaagc agcaagcaac
     3301 tttgactgct gtcttggata cacagaccgt attcttcatc ctaaatttat tgtgggcttc
     3361 acacggcagc tggccaatga aggctgtgac atcaatgcta tcatctttca cacaaagaaa
     3421 aagttgtctg tgtgcgcaaa tccaaaacag acttgggtga aatatattgt gcgtctcctc
     3481 agtaaaaaag tcaagaacat gggtacctag tgattagccg ctcgaggagg gcccgaacaa
     3541 aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcataa tcatcattga
     3601 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3661 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3721 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3781 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggacgcggcc
     3841 gcaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3961 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4021 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4081 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4141 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4201 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat gaccgagtac
     4261 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4321 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4381 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4441 tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg
     4501 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4561 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4621 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4681 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4741 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4801 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccgccac gaacttctct
     4861 ctgttaaagc aagcaggtga cgtggaagaa aaccccggtc ccggatccat ggtgtctaag
     4921 ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     4981 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     5041 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     5101 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     5161 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga ggacgggggc
     5221 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     5281 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     5341 gaggccttca ccgaaacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     5401 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
     5461 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     5521 agaatcaagg aggccaacaa cgaaacctac gtcgagcagc acgaggtggc agtggccaga
     5581 tactgcgacc tccctagcaa actggggcac aaacttaatt aagtcgacaa tcaacctctg
     5641 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5701 tgtggatacg ctgctttaat gcctttgtat catgcgttaa ctaaacttgt ttattgcagc
     5761 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5821 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggaattg
     5881 actcaaatga tgtcaattag tctatcagaa gctcatctgg tctcccttcc gggggacaag
     5941 acatccctgt ttaatattta aacagcagtg ttcccaaact gggttcttat atcccttgct
     6001 ctggtcaacc aggttgcagg gtttcctgtc ctcacaggaa cgaagtccct aaagaaacag
     6061 tggcagccag gtttagcccc ggaattgact ggattccttt tttagggccc attggtatgg
     6121 ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga agcgttcaga
     6181 ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc tgccggctcg
     6241 gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg ctgcccccta
     6301 gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc cctgatatct
     6361 ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa ataacaatat
     6421 aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg tcattttgac
     6481 tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag cacgcctcac
     6541 gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc gcgctattta
     6601 gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact atctttctag
     6661 ggttaatcta gctgcatcag gatcatatcg tcgggtcttt tttccggctc agtcatcgcc
     6721 caagctggcg ctatctgggc atcggggagg aagaagcccg tgccttttcc cgcgaggttg
     6781 aagcggcatg gaaagagttt gccgaggatg actgctgctg cattgacgtt gagcgaaaac
     6841 gcacgtttac catgatgatt cgggaaggtg tggccatgca cgcctttaac ggtgaactgt
     6901 tcgttcaggc cacctgggat accagttcgt cgcggctttt ccggacacag ttccggatgg
     6961 tcagcccgaa gcgcatcagc aacccgaaca ataccggcga cagccggaac tgccgtgccg
     7021 gtgtgcagat taatgacagc ggtgcggcgc tgggatatta cgtcagcgag gacgggtatc
     7081 ctggctggat gccgcagaaa tggacatgga taccccgtga gttacccggc gggcgcgctt
     7141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7321 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7381 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7441 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7621 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caa
//
