LOCUS       sequence-395158-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express kLight1.3 biosensor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  AUTHORS   Dong C, Gowrishankar R, Jin Y, He XJ, Gupta A, Wang H, Sayar-Atasoy 
            N, Flores RJ, Mahe K, Tjahjono N, Liang R, Marley A, Or Mizuno G, Lo
            DK, Sun Q, Whistler JL, Li B, Gomes I, Von Zastrow M, Tejeda HA, 
            Atasoy D, Devi LA, Bruchas MR, Banghart MR, Tian L
  TITLE     Unlocking opioid neuropeptide dynamics with genetically encoded 
            biosensors.
  JOURNAL   Nat Neurosci. 2024 Jul 15. doi: 10.1038/s41593-024-01697-1.
  PUBMED    39009835
REFERENCE   2  (bases 1 to 6217)
  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 
            Neurosci. 2024 Jul 15. doi: 10.1038/s41593-024-01697-1."
COMMENT     Sequence Label: pCMV- kappa-Light1.3
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        167..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1811..1832
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1938..1959)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2199..2218)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3111..3232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3202..3221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3239..3694)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3397)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3588..3607
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3721..3825
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3827..4184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3827..4018
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3850..3870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4035..4170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4219..5013
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4219..5013
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4273..4292)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4883..4902
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5201..5220)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5245..5292
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5621..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
       61 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
      121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      721 cgagctcaag cttaccggtg cgcgccatga agacgatcat cgccctgagc tacatcttct
      781 gcctggtgtt cgccgactac aaggacgatg atgacgccat ggattcacca attcagatat
      841 ttaggggtga accgggccct acctgcgcgc caagtgcttg tcttcctccg aattcaagcg
      901 catggttccc cggctgggca gagcccgact ctaatgggtc tgctggcagt gaagacgcgc
      961 agcttgaacc tgcccatatt agccctgcta tacccgttat catcaccgcc gtatattccg
     1021 ttgtattcgt ggtcggtctg gtaggaaatt ctctggtgat gtttgttatc atacgatata
     1081 ccaagatgaa aacagctacc aacatctaca tcttcaatct ggccttggcg gatgcgctgg
     1141 tcactactac tatgccattt caatcaactg tgtaccttat gaactcctgg ccttttggcg
     1201 atgtattgtg caagatcgtc atctcaatag attactataa tatgtttact tccatattta
     1261 ctctcactat gatgagcgtc gatcgatata tagcagtatg tcacccaaag aaggctttgg
     1321 actttcgaac gccattgaag gcgaaaatta taaacatttg catttggctg ctttcctcca
     1381 gcgttggaat ctccgctatt gtgttggggg ggaccaaggt cagagaagac gtagacgtga
     1441 tcgagtgttc cctgcagttc ccggacgatg attatagctg gtgggatctc tttatgaaga
     1501 tctgtgtctt catctttgct ttcgtaatac ctgtccttat cattattgta tgctatacgc
     1561 tgatgatact tagattgaaa tccgttagac tgagctcagg cattaacgtc tatatcaagg
     1621 ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg
     1681 gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1741 tgctgcccga caaccactac ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga
     1801 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1861 acgagctgta caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca
     1921 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1981 tgtccggcga gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2041 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2101 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2161 ccgaaggcta catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2221 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2281 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaaccca catgaccaac
     2341 tgagagagaa ggaccggaat ctgcgacgca ttaccagact ggtccttgtg gtcgtggcag
     2401 tgttcgtggt ctgctggact cccattcata tctttatact ggtggaggca ctcggttcta
     2461 cgagccatag cactgcggca cttgcagctt actacttctg tattgcactt gggtacacga
     2521 atgctgcact gaatcctatc ctgtacgcct tccttgatga gaatttcaaa agatgtttcc
     2581 gcgatttctg tttcccgttg aagatgcgga tggaaagaca agctaaggcc cgcgttagaa
     2641 atactgttca agaccctgca tatcttcggg acatagatgg tatgaataaa cccgttcagt
     2701 ctcagcccat cctgaacact aaggagatgg cccctcagag taaaccccct gaggaactgg
     2761 aaatgagctc catgccatct ccagtggctc ctctgccagc taggaccgag ggcgtgattg
     2821 acatgagaag catgtctagt atcgatagct tcatttcctg cgccaccgac ttccccgaag
     2881 ctacaaggtt tggaagcggt agcggttcag gatcaggctc tgcagagtca cctgcagatg
     2941 gcgaggtttt cgctatccag ctgacatctt cctgacaacc tgatgtacag cggccgcgac
     3001 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3061 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3241 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3301 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3361 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3421 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3481 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3541 cgatttagag cttgacgggg aaagtcggcg aacgtggcga gaaaggaagg gaagaaagcg
     3601 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3661 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3721 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3781 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3841 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4021 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4081 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4141 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4201 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4261 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4321 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4381 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4441 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4501 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4561 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4621 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4681 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4741 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4801 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4861 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4921 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4981 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5041 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5101 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5161 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5221 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5281 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5341 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5401 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5461 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5521 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5761 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccag
//
