LOCUS       sequence-466407-        7243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCR6-V2-PCP with a 4 amino acid linker and V2 tail.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7243)
  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 7243)
  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: LSM052 CMV-TO-CCR6-PCP (FLP-IN)
FEATURES             Location/Qualifiers
     source          1..7243
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      224..812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     713..732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     966..983
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1100..1121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1136..1141,1142..1159,1160..1166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1174..1190
                     /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     1179..1201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1198..1214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1198..1214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1263..1384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1274..1293)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1328..1347
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1514..2536)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(1514..2536)
                     /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(2544..2591)
                     /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)."
     primer_bind     2723..2742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2914..3138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3127..3144
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3200..3229)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5412..5436)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5446..5464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5467..5485
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5487..5690)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5516..5536)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5691..6070
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6237..6256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6331..6435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6436..6504,6505..7243,1..53)
                     /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(6654..6673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
       61 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
      121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
      181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
      241 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      301 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      361 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      421 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      481 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      541 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      601 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      661 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      721 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      781 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     1021 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     1081 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     1141 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1201 gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgacggt atacagacat
     1261 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1321 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1381 agttggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg
     1441 cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat
     1501 ctcgtagtac gtgctattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat
     1561 cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg
     1621 tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag
     1681 ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga
     1741 gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc
     1801 gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt
     1861 attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt
     1921 ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt
     1981 cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt
     2041 ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc
     2101 atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa
     2161 gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca gcgggcagtt
     2221 cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca
     2281 ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa
     2341 agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga
     2401 catatccacg ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct
     2461 gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg
     2521 tgagttcagg ctttttggcc aaggaagttc ctatactttc tagagaatag gaacttcgga
     2581 ataggaactt ctaggtacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2641 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2701 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2761 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2821 agggcgcgtg gggatacccc ctagagcccc agctggttct ttccgcctca gaagccatag
     2881 agcccaccgc atccccagaa catcgataga tctccataga gcccaccgca tccccagcat
     2941 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     3001 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     3061 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     3121 ggcaactaga aggcacagtc gaggctgatc agcgggttta aactcaatga tgattatgat
     3181 gatggtcgac ggcgctattc agatcctctt ctgagatgag tttttgttcg ggccctcctc
     3241 gagcggctca cctgcccaaa ggtacaagat tgacaaccag gtcttccacc tgtgaagttg
     3301 caacgagtga tttggtcaaa tcgtaaaggc ttttgcgaga cgcctcggta ctgttcgcca
     3361 caatggtcac gtcatgggac cacacctgcg tgtaccgtac ttttggcaga ccggaatcca
     3421 caacatccgc ttgatcgagt ttaaggttga ctcgataggc agttttggct ccgttctgtc
     3481 gcaatgaagc cgtcagccgc agccgcccga caagcggacc cactttctcc tcaaaaatct
     3541 gtctgtctgc tgttgactgg atttctgtca gtgtgcgggt tgcctctccc acgctcaaca
     3601 cgatggtctt actggcaagg gtacccccgc cggaaccgcc tccccgacca agaggcacca
     3661 aattgacgac aaggtcctca acttgagagg tggcgacgag tgacttagtg agatcataaa
     3721 ggcttttcct actcgcttcc gttgagtttg caacgattgt cacatcatgt gaccaaactt
     3781 gagtatatcg tactttgggc aagccgctat caacaacatc tgcttgatca agtttgagat
     3841 tcaccctata agcggtctta gcgccattct gccgcaggga agctgtcaac ctgagtcttc
     3901 cgacaagcgg tcccactttc tcttcgaaaa tttgtctatc cgctgtgctt tggatctccg
     3961 tgagggttct ggtggcttct cctacagaca acactattgt tttagaagct aggccggatc
     4021 cggtcgatga agtgtccttg gccagggagg agctggcggt ggtgcaggac tcatcttggg
     4081 gacccaggct gggtggggtg cgtccaccgg tcatggtaaa ggaggaggcg ttatcgttat
     4141 cggccgtctc actggtctgt ctggaaatgt tttcggagta gcgtcctgcg catgaaaatc
     4201 cactactttt atactttctt cttacacacc agaggtcttt gagaatcttc aggaagtagt
     4261 ttcgaaactt ctgacctata aatgcgtaca gcacagggtt aaggcaacaa tgcaggaacg
     4321 ccagaacctc cgtaaccgtt ttggtgtacc cgataagctt ttctgactga catgagcgat
     4381 tcattttgcc caggttggct gccgttacca gaaggaccat attatgaggg atctggcaag
     4441 ccagaaatac cagcacaaca gcgatgatta cccggatggc cttgtgtcgc ttagagtttt
     4501 gtgcctggac cagggtcttc acgatgaagg tataacaaaa aatcataaac atgagtggga
     4561 taaaaaagcc gaacagcaat tccaaaccca gcatcaacag cttccacctg ataggctcag
     4621 ataccgtctg gtacttgggc tcacacacat cgctgccctg cgtgttgtac ttctggttaa
     4681 acacgaaagt actgctggaa ataatcacgc tcaagcccca tacaacaaga caaatgattt
     4741 tagacctagg gagcgttcga gacctcagtc tgaaggattt tgttgcctgt actatggcga
     4801 tgtatctgtc catacttatg caagtcagca gcagcatacc gcaattaaag ttgatggcat
     4861 aaattccctt caacagcttg caggttgcat tactgaacac ccaggccccg gtagcgtgac
     4921 tcacagccca gaatgggagc gtcaggacga acaggatgtc tgcaatggcc atattaagca
     4981 ggtagacgtc tgtcatggat ctggcctttt tataaaaagc aaatgtgatg actaccaaga
     5041 tattccccag gaggccaaag acgcagatga gtgaatacgc gatgggaacg aagagtcggg
     5101 agaactggcg cacctcctgc agggagcaaa gcagcatctc agaatctacg ctgtagtagg
     5161 atgtattgac ggacacgaag tagtcctcac tggaatcgaa cacatcactg aagttcatgc
     5221 tctctccgct catatcgatg ctggcgtcat catcgtcctt gtagtcggcg aataccaggc
     5281 agaagatgta gctcagggcg atgatcgtct tcatggtggc gaagcttaag tttaaacgct
     5341 agagtccgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
     5401 cgtctccagg cgatctgacg gttcactaaa cgagctcgtc gacgatctct atcactgata
     5461 gggagatctc tatcactgat agggagagct ctgcttatat agacctccca ccgtacacgc
     5521 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     5581 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     5641 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     5701 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     5761 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     5821 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     5881 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     5941 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     6001 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     6061 aatcaatgtc aacgcgtata tctggcccgt acgaagcagc gcaaaacgcc taaccctaag
     6121 cagattcttc atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc
     6181 gcactactca gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc
     6241 atcagagcag attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg
     6301 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6361 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6421 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6481 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6541 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6601 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6661 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6721 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6781 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6841 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6901 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6961 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7021 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7081 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7141 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7201 atcgtagtta tctacacgac ggggagtcag gcaactatgg atg
//
