LOCUS       sequence-368449-        7995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mcm-3 under its native promoter (embryonic nuclei and 
            ciliated neurons IL2, CEMs, HOB, RnBs in C. elegans).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7995)
  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 7995)
  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: pIN2_mcm-3_mScarlet
FEATURES             Location/Qualifiers
     source          1..7995
                     /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(6555..6577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6677..6696
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6774..6792)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6860..6964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6965..7033,7034..7825)
                     /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(7183..7202)
                     /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 aaaatctgaa
     1261 tactacagta ctccttaaag gtgcacactc gtttgaagaa tagtaagcta atagattttg
     1321 tcttaaaaaa gtcgtttcga gacccggtac agtactcttg gaggcaacac gttcaaaatt
     1381 tcgccagtcc tgttataaat ctagattcaa ttctcacctg tctctgaaat ccagagtacc
     1441 agaatcgaga ttgcgagtag aagtttcatg gtttcgtatt cgtatcgtat tgcgaatata
     1501 aaacgaaaaa gcacagaaaa aatagctttg tgaatgaatg aatgagtgaa ggcccgagcg
     1561 gcggcgggag ggtccgatag atgttgtttc aaacggtttc caatggtttc cagcacgcgg
     1621 tgcgctcaag tttctaggag acaaagtcaa agcacttaaa gctttacggc ataaattttg
     1681 tgtatatgtg tgcttttgtg accactttct tcccatttct atgtataaat gtattacgaa
     1741 aggtgcgtcc tgggggaggc agggagaaaa taagaaagat gggatgaata gaatggaaaa
     1801 gcatctgtgt ctgtgagttt gtgagtactg aaacaataga ttccattttt gtgatttgtt
     1861 atgaaaaatt gaggagattt ttggaagaga tcttttgcat agatcacggg cacaaatgtt
     1921 attgatgttt tttgaaactt gaaatgacaa ttttgaactg ataatttcag acaattttca
     1981 ttttattcca ggcattttta tttaaataaa attatttcgc tattaatagc tatttaaaat
     2041 aaacatgttc aaacatcact ttcaggtgca ttgagtgcag cgttgcgtca tggtgtttac
     2101 ggagcgagcg cgcgaaacct tgctgtttga atgagaaatg cgcgaatttg ggccgaattt
     2161 ttctattttc cccgctcttc tccgtgcttt ctagtttcta ttcgtttatt ttttaatttt
     2221 aagtttcttt tgttaaaaac atcttatttt cagaaaaaat ggacggacaa gctgatggaa
     2281 tgttcattga tcgtgccgag gatgctcgaa aacaagaaat cacggagcag tatctcaact
     2341 ttttggacaa tcaagccgac acttttcact atcgtcagaa aattgtgaaa atgatgaatg
     2401 atggagaaag tcgtttggtg atgaatctcg atgaaatcag ggaaattcta ccagaaagag
     2461 ctgaagggta attaataaaa aataaagaat tcccgagtat taaagttttc aggctattga
     2521 cccaaagcgt tcttgagatg aattgcttac aatctgcact tcaaatggct attaatcgta
     2581 gtgatgttca agctgttgtc aaaacgtgag tagcttttaa aagccgtttt tgtttttaaa
     2641 caatttgtat tttcagtgga ttccacgtcg gctttgaagg aactttcgga gagcgccacg
     2701 tcaatccacg tactctcaaa tcatcatatc ttggaaacct tgtctgctgt gagggaatcg
     2761 tcacaaagtg ttcaactgtt cgtcaaaaac ttctcacttc tgtccactat tgtccagcaa
     2821 caaataaagt gctcgagaag aaatttgccg atttcactat gctcgacact gcagtcacca
     2881 ataacaacta tccaacagaa gatgaaaata agaatcctct tgaaacagaa ttcggacatt
     2941 cagtttacaa ggatcatcaa acgttcacaa ttcaagagct tccagaatcc gcaccagccg
     3001 gtcaacttcc tcgggccgtc gattgtgttg ctgatcttga tttggcagat cgcgtcaagc
     3061 ctggagatcg tgtccgcatc attggagtct tccgtgttct gccgaataag cagaatggag
     3121 tttcgtctgg atccttccgt tcgatcatca tcattaatca tattcaaatg ctttcaaagg
     3181 aggttattcc aaactttgag ccacaagatg ttaaagatgt tcgtaaaatt tcaaagtcaa
     3241 gaaacccatt tgagctgctc gctcggtcac ttgctccatc aatttgcggc cacgaagaga
     3301 caaagaaagc gttgctctgt ttgttgctag gtggaatgga aaagattctg aacaatggtt
     3361 ctcgtttgag gggagatatc aatgttcttc tcatcggaga tccatctgtt gccaagtctc
     3421 aactgcttcg atatgttctt cgtatggccc caagagctat caccacaact ggacgtggat
     3481 cttcgggtgt tggacttact gctgctgtta ctactgatcc tgattcggga gaacgtcgtc
     3541 ttgaagctgg agcaatggtt ctcgccgatc gtggtgtcgt ttgtattgat gaatttgaca
     3601 agatgtctga tattgataga actgcaatcc acgaagttat ggaacaggga cgtgtcacaa
     3661 tttcgaaggc tggaatccac gcgaagctga atgctcgttg ctcggttctg gctgcagcaa
     3721 atccggtgta cggacgatac aatccattca aatctccaat ggaaaacatc ggaatgcaag
     3781 attcactcct ctcgcgtttc gatttgatct ttgtacttct tgatgagcat gatgcggaca
     3841 aggacgctaa tgtcgctgaa catgtgctca agcttcacac ttacagaacc caaggtgaag
     3901 ccgatggaac tgttcttccg atgggtggag gtgttgagac gatcagtaca atcaatatgg
     3961 agactaagaa ggcgagcagc tcgatctatg aggaaaatac ccaatgggct ggtattcaag
     4021 agtgagtttc ttttataaaa aaacaatata aattaattaa tttctctatt tcagcacgaa
     4081 aatcctgaca atggacttca tgagaaagta catccatttg gcaagatcag ttcaaccaaa
     4141 actcaccgat gaagctactg agtacatttc tgaagtatat gccgatattc gttcatttga
     4201 tatcaccaaa actgatcaag aacgaacaat gcctgtcacc gctcgtcaac tcgaaacatt
     4261 gattcgtttg tcaactgcta tcgcgaaggc aagattctcg aaaaaagtcg aacgagagga
     4321 tactgagaaa gcttacaatc tgctccactt tgcctgtttc aaggagaagc caaaggctcg
     4381 tcaagagtat gagaagaaga agcgagctcc acgaattgat ggtgacgatg atgaggatca
     4441 gagcgataac gaggacatgg caacatcgga agagcctcaa tctagctccg ttccgactgc
     4501 agccaccccg cgtcgtggag tcagaaggcg tgctgaagat gatgatagcc aggattctat
     4561 gatggatact accattggag aaagtcagcc aactgctaag aaagcacgta ctggaccggc
     4621 tgccatcgga gttgatcgtt ataaggagct tcggaagtac gtcagaaagg cgttcgacga
     4681 cattggaagc accgacgata tggttgaact tcaaattatc acagacagta agcaaatgtt
     4741 tcattaaaaa tgtttattac aaaaatgttt tcaggtgttc aaactcaagc cggatcacac
     4801 aagttcactg aggatgagct gaaggctgga tacgaacaat tggaaaacga taacgctgca
     4861 atgattgccg aagataagat cactctcatc ggaggtggcg gatctggagg tggaggctct
     4921 ggaggaggtg gatctgtcag caagggagag gcagttatca aggagttcat gcgtttcaag
     4981 gtccacatgg agggatccat gaacggacac gagttcgaga tcgagggaga gggagaggga
     5041 cgtccatacg agggaaccca aaccgccaag ctcaaggtca ccaagggagg accactccca
     5101 ttctcctggg acatcctctc cccacaattc atgtacggat cccgtgcctt catcaagcac
     5161 ccagccgaca tcccagacta ctacaagcaa tccttcccag agggattcaa gtgggagcgt
     5221 gtcatgaact tcgaggacgg aggagccgtc accgtcaccc aagacacctc cctcgaggac
     5281 ggaaccctca tctacaaggt caagctccgt ggaaccaact tcccaccaga cggaccagtc
     5341 atgcaaaaga agaccatggg atgggaggcc tccaccgagc gtctctaccc agaggacgga
     5401 gtcctcaagg gagacatcaa gatggccctc cgtctcaagg acggaggacg ttacctcgcc
     5461 gacttcaaga ccacctacaa ggccaagaag ccagtccaaa tgccaggagc ctacaacgtc
     5521 gaccgtaagc tcgacatcac ctcccacaac gaggactaca ccgtcgtcga gcaatacgag
     5581 cgttccgagg gacgtcactc caccggagga atggacgagc tctacaagta gcattcgtag
     5641 aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg
     5701 ggccgctgtc atcagagtaa gtttaaactg agttctacta actaacgagt aatatttaaa
     5761 ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt caacatccct
     5821 acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa aaacttctct
     5881 taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc
     5941 aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa
     6001 taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttactt
     6061 ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt
     6121 tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt
     6181 caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa
     6241 agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt gtttgtcaag
     6301 atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa
     6361 agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt tgataatttg
     6421 aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt cccaatatac
     6481 caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt
     6541 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6601 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6661 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6721 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta agggcctcgt
     6781 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6961 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7021 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7081 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7141 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7201 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7261 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7321 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7381 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7441 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7501 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7561 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7621 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7681 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7741 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7801 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7861 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7921 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7981 gatcaaagga tcttc
//
