LOCUS       sequence-368447-        7953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tsp-6::mScarlet under its native promoter (amphid and 
            phasmid ciliated neurons in C. elegans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7953)
  AUTHORS   Nikonorova IA, Wang J, Cope AL, Tilton PE, Power KM, Walsh JD, 
            Akella JS, Krauchunas AR, Shah P, Barr MM
  TITLE     Isolation, profiling, and tracking of extracellular vesicle cargo in
            Caenorhabditis elegans.
  JOURNAL   Curr Biol. 2022 May 9;32(9):1924-1936.e6. doi: 
            10.1016/j.cub.2022.03.005. Epub 2022 Mar 24.
  PUBMED    35334227
REFERENCE   2  (bases 1 to 7953)
  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"; doi: 
            "10.1016/j.cub.2022.03.005"; journalName: "Curr Biol"; date: 
            "2022-05-9- 9"; volume: "32"; issue: "9"; pages: "1924-1936.e6"
COMMENT     Sequence Label: pIN4_tsp-6_mScarlet
FEATURES             Location/Qualifiers
     source          1..7953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6513..6535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6635..6654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6732..6750)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6818..6922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6923..6991,6992..7783)
                     /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(7141..7160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgt aagtttaaac
     1021 atgatcttac taactaacta ttctcattta aattttcaga gcttaaaaat ggctgaaatc
     1081 actcacaacg atggatacgc taacaacttg gaaatgaaat aagcttgcat gcctgcaggt
     1141 cgactctaga ggatccccgg gattggccaa aggacccaaa ggtatgtttc gaatgatact
     1201 aacataacat agaacatttt caggaggacc cttggagggt accggtagaa aaagtttttg
     1261 tcactttttt ggatcaaaat tttgttcaat tgcctattta tttgcaaaat acccaggttc
     1321 aaaattggta tgggttcgaa catgtgacag tagggcttcc atggcgtcag gccctccagg
     1381 gccccacgca ggccaccctg caggccattt aggcagggaa gcctcatgcg acaaggcttt
     1441 gtttttttca aaatcggcca aaaatttagt aattgtcact tacacgtttt ttcccaataa
     1501 aaaattttcc attaaaaaat tttgcgaaat ttttgcatag tttcatatgt agttgatcat
     1561 ttcgacatta aagaagtgat gctttcttat tcagtttcag tttcttaaaa cttctcaaaa
     1621 tcaaaaaagt gtttcttctt tcattccaac tttgtccatg attcatactg aaaaaatttt
     1681 aaaaaatgtt ttgccaacac cttttgttgg ttttgttaca gaatatcaaa acaggaactg
     1741 gtacacaaac atgtttttat tttctttaat cagaaaaacg aggcgttatc aactttgtcg
     1801 cttggagttt tgaaatttac cagcttgtct ccaattttaa aactaaacta ctacaaatag
     1861 agcttgacgt gggttgcaga cgtcatttaa tatcattcta tgattccgtt tactcatgag
     1921 attccagcta atttacactt tcgatgaagc aagtacgcac tctccatttc ctgtgattgt
     1981 gcataactgg cgtcaaggta accgatgcac acaatacacc ggtgcctctt agcccgcccg
     2041 acttctgatg tgaatcttgc attcgagaag cagcactggc ggcggtcgac atgtacatat
     2101 caccagtctg gcttttcccg tttccgcgag caagcggcgg gtggttgttg ttatgggata
     2161 ccggagccaa ttgatagcaa atctccggtt gctattcgtt gcgcactttg attacttttt
     2221 agtgcatttt tctgcttgct tccgattgta ttcttccgtt tttttttcat ttagaattgt
     2281 gatcttgcgc acttttgaat tgaaattacc acaaatttat gtatttctaa tcctaacaca
     2341 tttttatcaa agctttgagc tgtgttttta tgtcttgtga tccaacagaa taaaaatgac
     2401 caaacgtcaa attaatatta ttcaaaaaca ttttggaaat gtttttattc actgtcaaaa
     2461 agtgacaatt aatcagttct taatatttga gttcgaccaa aaaaaaaaac actcctacga
     2521 tttgtgttaa aacatgcgag ggtcggaaca tgctacaatg ctctaaattt ccttaaaagc
     2581 tcttttcata tatttcggca atattccata acgttgtctg gaaattgaag gtgtttacac
     2641 aaaatcccca ctggcgctac tccacctgta accatataat ttaaatcagc tctctcacgc
     2701 gcattgctgc gtatttcgta attctgatag cataatcata accagaatac gaaaaactgc
     2761 ttgtccctcc cacagtagtc actcacatga atgaaattat cagaatatca ctcgtgagct
     2821 ttcacttttt tagcatgtct gttttctaac tacacgagca gctgttttat ttaaattggc
     2881 catgggaggt taagaaaaca aatgaatagg tggccataaa acgtgatatt taagagctcc
     2941 tgaagccaca aatttcccac ggagaaagcg aagagttagc tggcaaaggg gtacatactt
     3001 gagcagtagg atgaccaatt agcatttctc gtgcgctctc aatcttgttc tacttgctta
     3061 atatcgcttt tcgggttctt cctttttctg cgctgttgct cctttttcac gttttccttc
     3121 ttctttttct tcttcatttt ttcttacccg ttgtctcttg ttgctcctct ttttcacatt
     3181 cacattggtc gttgttcgtt tcattttata tcatcatccg gttgtttctt gttttcaatc
     3241 ttgctttttt tccggcctct cgtcttccaa gatggttcaa ggatgtggta acaaatgcgt
     3301 caagtacttt ttctggctca tcaactttct gttctttgta agttttaata aatttgagta
     3361 ttttaacgta caagccgaaa atgtgctcaa ataaaccaat tttgtaatta agattttcaa
     3421 aaatttttta aattttttta tagttatggg cacaaaaaat tgtcctagtc tcaaaattgt
     3481 cctagtctct tttgtcctag tcgctaaaat ttcgaatttt gcacaatttt ggtggaactt
     3541 atcttttttt tgaattacaa tccttcttct ttgcatgttt tagttgttta tctcaccatt
     3601 tttttaaata cagtagaccg aaagtcgagg tatccaaaaa tttccaaaat tggctactta
     3661 tcagcattgt accttaggca acgaaataac gtgagataaa cgacaaaata tgcaacaaag
     3721 aatgataatt caaagaaatt gttaattttt tttttgaaaa aaaagggaaa aaagtataaa
     3781 aaattggctg aaagttggta tttgccgaaa taattttttt tggcaaatgg caacttttaa
     3841 tttttatttt aaagtaataa aatatttact tctacttcta catttagtct tattttaaaa
     3901 atgggttatc atatcggaaa aatcagtttc gttgaaacac attttgcgtt tatacgtcac
     3961 cataaccgtt actcctgttt taatttatat actattttca tgttcttggg gcaataatcg
     4021 tcgggctctc catatggatg ctagtggaca agaattctct gaatacggtg gcaagtacag
     4081 tgaaagttga tctctcccaa attttgagtc aagtgaatat tcaacaggtg attcaaccat
     4141 aattttaata ataacttgtg agtcaaaatc tacagctcaa ctcgttcctc tacgttgcga
     4201 tcgtaatcgg aggggctctt ctggttctcg gattcttcgg ttgctgtgga agttgctgcg
     4261 aaagcatctg tgctatttcc atctacttca tcttggtcct catcctcttc gttgttgaag
     4321 tcgtcgccat cgttctctac tttgttaaca agactaacgt aatgtgacgc cttccagatg
     4381 ctctcataaa ttccatgttt cagcttcaac aagggttcca aactatttgg cgcgacgaac
     4441 tggtttcaaa gtataacact caacaacaga ttcaccaagt gctcgaccaa atccaatcta
     4501 gcgtgagtta taatattttt agattttcac aagatcagct tcaatcaaag tatgcaattt
     4561 tagcttcaat gctgtggagc ttctggatgc agtgactaca taccctacgg agcattccca
     4621 acctcctgcc aatgtgcaac tatccaacaa gtaacttgta aagcatttga gctcagactc
     4681 taaccgttcc aaatttaggc tggatgcgct actgttatct ggaactcatt tgaaagcagc
     4741 ctgatttacg tcgccttcgt tggaatcatc atcctttttg tcgagctgct tgcgatgata
     4801 ttctcttgta tcattattgg ggccgtcaaa gagaaacgct cccaagctgg aggtggcgga
     4861 tctggaggtg gaggctctgg aggaggtgga tctgtcagca agggagaggc agttatcaag
     4921 gagttcatgc gtttcaaggt ccacatggag ggatccatga acggacacga gttcgagatc
     4981 gagggagagg gagagggacg tccatacgag ggaacccaaa ccgccaagct caaggtcacc
     5041 aagggaggac cactcccatt ctcctgggac atcctctccc cacaattcat gtacggatcc
     5101 cgtgccttca tcaagcaccc agccgacatc ccagactact acaagcaatc cttcccagag
     5161 ggattcaagt gggagcgtgt catgaacttc gaggacggag gagccgtcac cgtcacccaa
     5221 gacacctccc tcgaggacgg aaccctcatc tacaaggtca agctccgtgg aaccaacttc
     5281 ccaccagacg gaccagtcat gcaaaagaag accatgggat gggaggcctc caccgagcgt
     5341 ctctacccag aggacggagt cctcaaggga gacatcaaga tggccctccg tctcaaggac
     5401 ggaggacgtt acctcgccga cttcaagacc acctacaagg ccaagaagcc agtccaaatg
     5461 ccaggagcct acaacgtcga ccgtaagctc gacatcacct cccacaacga ggactacacc
     5521 gtcgtcgagc aatacgagcg ttccgaggga cgtcactcca ccggaggaat ggacgagctc
     5581 tacaagtagc attcgtagaa ttccaactga gcgccggtcg ctaccattac caacttgtct
     5641 ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt ttaaactgag ttctactaac
     5701 taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg cctctgactt ctaagtccaa
     5761 ttactcttca acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta
     5821 ttatcaaaaa acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca
     5881 atgaaattgt gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt
     5941 gctccctccc cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac
     6001 ttcttatgtt ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca
     6061 aaatacctta tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt
     6121 ctgggcctct catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga
     6181 atttttcaat caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt
     6241 cccctattgt ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg
     6301 atgagcacga tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga
     6361 gtagaagttg ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag
     6421 aatacattcc caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt
     6481 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     6541 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     6601 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     6661 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     6721 cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6781 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6841 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6901 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6961 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7021 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7081 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7141 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7201 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7261 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7321 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7381 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7441 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7501 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7561 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7621 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7681 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7741 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7801 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7861 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7921 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
