LOCUS       sequence-478944-        5534 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses CCR6-NanoLuc in mammalian cells; used with 
            LATeNT*-Arrestin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5534)
  AUTHORS   Roh H, Kim D, Kim B, Jeon Y, Kim Y, Jacko M, Xu F, Lin C, Won Um J, 
            Ting AY
  TITLE     Light-activated tetanus neurotoxin for conditional proteolysis and 
            inducible synaptic inhibition in vivo
  JOURNAL   bioRxiv 2025.01.27.635161; doi: 10.1101/2025.01.27.635161
REFERENCE   2  (bases 1 to 5534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.01.27.635161; doi: 10.1101/2025.01.27.635161"
COMMENT     Sequence Label: pAAV-CCR6-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1967..2476
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             2477..2518
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2555..2572)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2561..2768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2797..2937
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2797..2926
                     /label=AAV2 ITR
     rep_origin      3012..3467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3099..3118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3309..3330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3484..3503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3603..3625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3663..3681)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3749..3853
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3854..3922,3923..4714)
                     /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(4072..4091)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4885..5473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5374..5393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgagcggag agagcatgaa cttcagtgat gtgttcgatt ccagtgagga
      841 ctacttcgtg tccgtcaata catcctacta cagcgtagat tctgagatgc tgctttgctc
      901 cctgcaggag gtgcgccagt tctcccgact cttcgttccc atcgcgtatt cactcatctg
      961 cgtctttggc ctcctgggga atatcttggt agtcatcaca tttgcttttt ataaaaaggc
     1021 cagatccatg acagacgtct acctgcttaa tatggccatt gcagacatcc tgttcgtcct
     1081 gacgctccca ttctgggctg tgagtcacgc taccggggcc tgggtgttca gtaatgcaac
     1141 ctgcaagctg ttgaagggaa tttatgccat caactttaat tgcggtatgc tgctgctgac
     1201 ttgcataagt atggacagat acatcgccat agtacaggca acaaaatcct tcagactgag
     1261 gtctcgaacg ctccctaggt ctaaaatcat ttgtcttgtt gtatggggct tgagcgtgat
     1321 tatttccagc agtactttcg tgtttaacca gaagtacaac acgcagggca gcgatgtgtg
     1381 tgagcccaag taccagacgg tatctgagcc tatcaggtgg aagctgttga tgctgggttt
     1441 ggaattgctg ttcggctttt ttatcccact catgtttatg attttttgtt ataccttcat
     1501 cgtgaagacc ctggtccagg cacaaaactc taagcgacac aaggccatcc gggtaatcat
     1561 cgctgttgtg ctggtatttc tggcttgcca gatccctcat aatatggtcc ttctggtaac
     1621 ggcagccaac ctgggcaaaa tgaatcgctc atgtcagtca gaaaagctta tcgggtacac
     1681 caaaacggtt acggaggttc tggcgttcct gcattgttgc cttaaccctg tgctgtacgc
     1741 atttataggt cagaagtttc gaaactactt cctgaagatt ctcaaagacc tctggtgtgt
     1801 aagaagaaag tataaaagta gtggattttc atgcgcagga cgctactccg aaaacatttc
     1861 cagacagacc agtgagacgg ccgataacga taacgcctcc tcctttacca tgactagtga
     1921 gctggccgag aagctggccg gcctggacat caacggtgga gctagcgtct tcacactcga
     1981 agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca
     2041 gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga tccaaaggat
     2101 tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc cgtatgaagg
     2161 tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga
     2221 tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc
     2281 gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa
     2341 gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa
     2401 ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg
     2461 cgaacgcatt ctggcgggca agccgatccc taaccctctg ctgggcctgg acagcactta
     2521 gtaagaattc ctcgagctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     2581 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2641 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2701 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2761 gctggggagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     2821 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     2881 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     2941 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3001 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3061 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3361 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3481 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3541 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3601 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3661 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3721 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3781 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3841 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3901 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3961 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4021 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4081 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4141 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4201 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4261 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4321 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4381 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4441 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4501 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4561 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4621 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4681 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4741 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4801 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4861 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4921 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4981 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5041 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5101 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5161 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5221 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5281 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5341 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5401 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5461 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5521 ttttgctcac atgt
//
