LOCUS       sequence-478952-        8518 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses LATeNT*-Arrestin in mammalian cells; used with 
            CCR6-NanoLuc or KORD-NanoLuc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8518)
  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 8518)
  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: pcDNA3-LATeNT(slow)-Arrestin
FEATURES             Location/Qualifiers
     source          1..8518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4011..4040
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(4071..4089)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4072..4089)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4109..4126)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4115..4339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4385..4813
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4472..4491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4682..4703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4822..4842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4827..5156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5007..5142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5069..5088
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5223..6017
                     /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(5277..5296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5887..5906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6191..6312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6228..6247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6282..6301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6361..6377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6361..6377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6374..6396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6385..6401
                     /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(6409..6415,6416..6433,6434..6439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6454..6475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6592..6609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6763..7351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6843..6862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7522..8313,8314..8382))
                     /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     8145..8164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8383..8487)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gcatgccgat caccatcaac aacttccgtt actccgaccc ggttaacaac gacaccatca
      961 tcatgatgga accgccgtac tgcaaaggtc tagacatcta ctacagagcc ttcaaaatca
     1021 ccgaccgtat ctggatcgtt ccggaacgtt acgaatttgg taccaaaccg gaagacttca
     1081 acccgccgtc ctccctgatc gaaggtgctt ccgaatacta cgacccgaac tacctgcgta
     1141 ccgactccga caaagaccgt ttcctgcaga ccatggttaa actgttcaac cgtatcaaaa
     1201 acaacgttgc tggtgaagct ctgctggaca aaatcatcaa cgctatcccg tacctgggta
     1261 actcctactc cctgctggac aaattcgaca ccaactccaa ctccgtttcc ttcaacctgc
     1321 tggaacagga tccgtccggt gctaccacca aatccgctat gctgaccaac ctgatcatct
     1381 tcggtccggg tccggttctg aacaaaaacg aagttcgtgg tatcgttctg cgtgttgaca
     1441 acaaaaacta cttcccgtgc cgtgacggtt tcggttccat catgcagatg gctttctgcc
     1501 cggaatacgt accgaccttc gacaacgtta tcgaaaacat cacctccctg accatcggta
     1561 aatccaaata cttccaggac ccggctctgc tgctgatgca cgaactgatc cacgttctgc
     1621 acggtctgta cggcatgcag gtttcctccc acgaaatcat cccgtccaaa caggaaatct
     1681 acatgcagca cacctacccg atctccgctg aagagctctt caccttcggt ggtcaggacg
     1741 ctaacctgat ctccatcgat atcaaaaacg acctgtacga aaaaaccctg aacgactaca
     1801 aagctatcgc taacaaactg tcccaggtta cctcctgcaa cgacccgaac atcgacatcg
     1861 actcctacaa acagatctat cagcagaagt atcagtttga taaggattcc aacggccagt
     1921 atatagtgaa cgaggataag tttcagatac tgtataactc cataatgtat ggcgggagcg
     1981 gcagcggggg gtccggtagc ggggagtttc gggcaaccac actggaacgg atcgagaaaa
     2041 gcttcctgat tactgatccg agactgcctg acaacccaat catttttgtg agcaattcct
     2101 tcctgcagct gacagaatat tctcgggaag agatcctggg gcgcaattgc cgttttctgc
     2161 agggacccga gacagaccgt gccactgttc ggaaaatcag agatgctatt gacaaccaga
     2221 ctgaagtgac cgttcagctg atcaattata ccaagagcgg caagaagttc tggaacgtgt
     2281 tccacctgca gccgatgcgc gattataagg gcgacgtcca gtacttcatt ggcgtgcagc
     2341 tggatggcac cgaacgcctt catggcgccg ctgagcgtga ggcggtctgc ctggtcaaaa
     2401 agacagcctt tcagattttt actgagatag agctgggcaa gaagtttaac ataaagactc
     2461 gcttaagcta tttttccatg aaccacgatc ctgtgaagat acctaacctg ctggatgata
     2521 ctatatataa cgatactgag ggctttaaca tagagtccaa ggatctgaag tccgagtata
     2581 agggccagaa catgagggtg aacactaacg cctttaggaa cgtcgacggt tccggtctgg
     2641 tttccaaact gatcggtctg tgcaaaaaaa tcatcccgcc gaccaacatc cgtgaaaacc
     2701 tgtacaaccg taccgctggc tcgggcggga gtaccggtgg tagtgggagc ggtggtgaaa
     2761 aacccgggac cagggtcttc aagaagtcga gccctaactg caagctcacc gtgtacttgg
     2821 gcaagcgtga ctttgtggat cacttggaca aagtggatcc tgtcgatggt gtggtgcttg
     2881 tggatcctga ctacttgaag gaccggaaag tgtttgtgac cctcacctgt gccttccgct
     2941 atggccgaga agacctggat gtactgggcc tgtctttccg caaagatctg ttcatcgcca
     3001 cctaccaggc cttccccccc atgcccaacc cacctcggcc ccccacccgc ctacaggacc
     3061 gactgctgaa gaagttgggc cagcacgccc accccttttt tttcacaata ccccagaatt
     3121 tgccttgctc cgtcacactg cagccaggac cggaggacac agggaaggcc tgtggagtag
     3181 actttgagat tcgagccttc tgtgccaaat ctatagaaga aaaaagccac aaaaggaact
     3241 ccgtgcggct tatcatcaga aaggtacagt ttgctcctga gacacccggc ccccagccat
     3301 cagctgaaac cacacgccac ttcctcatgt ctgaccggag gtccctgcac ctagaggctt
     3361 ccctggacaa agagctgtac taccatgggg aacccctcaa tgtcaacgtc cacgtcacca
     3421 acaattctgc caaaaccgtc aagaagatca gagtgtctgt gagacagtat gccgacattt
     3481 gcctcttcag caccgcgcag tacaagtgtc ctgtggctca gcttgaacaa gatgaccagg
     3541 tgtctcccag ttccacattc tgcaaggtgt acaccataac cccgctgctc agtgacaacc
     3601 gagagaagcg tggccttgcc cttgatgggc aactcaagca cgaagacacc aacctggctt
     3661 ccagcaccat tgtgaaggag ggagccaaca aggaggtgct gggaatccta gtatcctaca
     3721 gggtcaaggt gaagctggtg gtgtctcgag gcggggatgt ctccgtggag ctacctttcg
     3781 tcctaatgca ccccaagccc cacgaccaca tcacccttcc ccgaccccag tcagcccccc
     3841 gggaaataga catccctgtg gataccaacc tcattgaatt cgataccaac tatgccacag
     3901 acgacgacat cgtgtttgag gactttgcga ggcttcggct gaaggggatg aaggatgacg
     3961 actgtgatga ccagttctgc ggtggaagtg gatcaggcag cggtggatct gaacaaaagc
     4021 ttatttctga agaggacttg taactcgagc atgcatctag agggccctat tctatagtgt
     4081 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4141 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4201 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4261 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4321 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     4381 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4441 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4501 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4561 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4621 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4681 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4741 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4801 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4921 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4981 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5041 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     5101 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5161 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     5221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5401 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5521 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6001 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6061 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6121 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6181 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6241 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6301 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     6361 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6421 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6601 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7021 caaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7201 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7441 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7501 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7561 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7621 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7681 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7741 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7801 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7861 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7921 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7981 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8041 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8101 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8161 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8221 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8281 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8341 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8401 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8461 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
