LOCUS       sequence-466396-        5467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  plasmid expressing secreted soluble CCL20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5467)
  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 5467)
  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: XZ034 CMV-TO-CCL20
FEATURES             Location/Qualifiers
     source          1..5467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    308..532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     521..538
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(594..623)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1030..1054)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1064..1082
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1085..1103
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(1105..1308)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1134..1154)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1309..1688
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1855..1874
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1949..2053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2054..2122,2123..2914)
                     /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(2272..2291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3085..3673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3574..3593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3827..3844
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3961..3982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3997..4002,4003..4020,4021..4027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4035..4051
                     /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     4040..4062
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4059..4075
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4059..4075
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4124..4245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4135..4154)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4189..4208
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4375..5397)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(4375..5397)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(5405..5452)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
       61 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      121 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      181 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtggggata ccccctagag
      241 ccccagctgg ttctttccgc ctcagaagcc atagagccca ccgcatcccc agaacatcga
      301 tagatctcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
      361 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
      421 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
      481 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
      541 gatcagcggg tttaaactca atgatgatta tgatgatggt cgacggcgct attcagatcc
      601 tcttctgaga tgagtttttg ttcgggccct cctcgagcgg cttacatgtt cttgactttt
      661 ttactgagga gacgcacaat atatttcacc caagtctgtt ttggatttgc gcacacagac
      721 aactttttct ttgtgtgaaa gatgatagca ttgatgtcac agccttcatt ggccagctgc
      781 cgtgtgaagc ccacaataaa tttaggatga agaatacggt ctgtgtatcc aagacagcag
      841 tcaaagttgc ttgctgcttc tgattcgccg cagaggtgga gtagcagcac tgacatcaaa
      901 gcagccagga gcaaactctt ggtacagcac atggtggcga agcttaagtt taaacgctag
      961 agtccggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg
     1021 tctccaggcg atctgacggt tcactaaacg agctcgtcga cgatctctat cactgatagg
     1081 gagatctcta tcactgatag ggagagctct gcttatatag acctcccacc gtacacgcct
     1141 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     1201 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     1261 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     1321 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     1381 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     1441 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     1501 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     1561 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     1621 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     1681 tcaatgtcaa cgcgtatatc tggcccgtac gaagcagcgc aaaacgccta accctaagca
     1741 gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc
     1801 actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat
     1861 cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac
     1921 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1981 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2041 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     2101 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     2161 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     2221 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     2281 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     2341 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2401 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2461 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2521 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2581 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2641 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2701 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2761 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2821 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2881 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2941 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     3001 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     3061 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3121 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3181 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3541 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3601 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3661 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3721 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3781 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3841 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3901 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3961 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     4021 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     4081 ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag
     4141 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4201 gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa
     4261 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     4321 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtagta cgtgctattc
     4381 ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca
     4441 gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc
     4501 tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc
     4561 gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg
     4621 cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca
     4681 agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat
     4741 cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga
     4801 gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag
     4861 ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg
     4921 atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat
     4981 tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc
     5041 atccatggcc tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg
     5101 caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc
     5161 aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg
     5221 atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac
     5281 atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct
     5341 gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttggc
     5401 caaggaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tctaggtacg
     5461 tgaacca
//
