LOCUS       sequence-315115-        5458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent co-injection marker to identify extrachromosomal arrays 
            in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5458)
  AUTHORS   El Mouridi S, Peng Y, Frokjaer-Jensen C
  TITLE     Characterizing a strong pan-muscular promoter (Pmlc-1) as a 
            fluorescent co-injection marker to select for single-copy 
            insertions.
  JOURNAL   MicroPubl Biol. 2020 Sep 3;2020. doi: 
            10.17912/micropub.biology.000302.
  PUBMED    32908967
REFERENCE   2  (bases 1 to 5458)
  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: "MicroPubl 
            Biol."; date: "2020-09-3"; volume: "2020. doi"; pages: " 
            10.17912/micropub.biology.000302"
COMMENT     Sequence Label: pSEM228 - [Pmlc-1 | mNeonGreen(2x NLS) | cbr-t...
FEATURES             Location/Qualifiers
     source          1..5458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..22)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5..21)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14..36)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     349..371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(409..427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        495..599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(600..668,669..1460)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(818..837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1631..2219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2120..2139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2346..2368
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2365..2381
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2365..2381
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2401..2421
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    3737..3761
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3770..3790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4135..4144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4690..4761
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(4769..4793)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5435..5455)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
       61 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      121 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
      181 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt gcaatctgct
      241 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
      301 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
      361 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
      421 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
      481 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      541 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      601 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      661 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      721 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      781 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      841 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      901 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      961 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1021 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1081 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1141 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1201 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1261 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggctctc
     1321 gcggtatcat tgcagcactg gggtcagatg gtaagccctc ccgtatcgta gttatctaca
     1381 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1441 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1501 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1561 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1621 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1681 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1741 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1801 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1861 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1921 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1981 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2041 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2101 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2161 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2221 acgccagcaa cgcggccttt ttacggttcc tggtcttttg ctggcctttt gctcacatgt
     2281 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2341 ataccagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
     2401 caactttgta tagaaaagtt gagcgtttct gccagctgac gcattatctt aaaagctgtt
     2461 ttcagataag agcactggta gacagaacac tcaatagaaa atcagatgaa aaagtttgaa
     2521 agttgaggaa tcaaaaaagt tatccttgtt aatttaggat atgagtaaga acctcttgtc
     2581 acactaattc tgatggaagg tttttttttc tgaatcgttt ctgaattgtc tccaatgaga
     2641 gtatagttcg aaacacaatt tttgtctgtt attcaatgac atcaaaatta ttgatttcaa
     2701 aaatagctgc aaatcctaaa atcaaaaatc actgcgtatg tcaaaacttt gaagattgtg
     2761 tgaaaatcaa tatttgctct gtttcatcct atcttaaaat acgcccgatt aattatttca
     2821 ttttaacaaa acttgaaaaa aaactataca aatagatttc acaagtagtt gaaacaatta
     2881 gtaggaaaat tataatttat accagaaaaa tatatgtcaa aaaaaattat cgttagtgaa
     2941 atattataaa cagatattac tggaaccatc tgataaattc agtatgtccc atttagtcat
     3001 gttatgaaaa ttaaaattta taatcaagtt ccaaaacaaa agagtttcag attttcaagt
     3061 tttcaattat cctgtttgtt ggaaatgttg aatttaaacc ttcctttcca actcaaaaca
     3121 aattctgatt atgttgtgcc ctgctcagat ttaaagctcg gatgatctat tccctttccc
     3181 ttattgcatc ccctcattta tccgatcgag caactttttc gatggcgggt aagaggcatc
     3241 gcgacgcaag caaggcgaga gttttgccca tcttcttgga aacctccgac gggtgcctga
     3301 tgttagcacc acgggggcgg tgcgcctctt gtgtaaagtt gttgagattt gctcaaacag
     3361 ccctcccctc gacgcggggc ccaacgagtg taaaaggcag acaaaagcga cggataatct
     3421 tcagttagtc tcagttcact actcagtaca gtccacaaaa tggtgagttg gattttttaa
     3481 taagttattt gaaaagcatg agtttctgta taggaatagg agtgcgttca tatatatttt
     3541 gatttaacct gtctatattg tatttaaaaa attatttatt ttgaatcaga ggttctataa
     3601 tcagtttcat ctttaaaaaa tttctggtaa attaagaacg taacatttag atttaggctt
     3661 tgtttaaaaa tcaagcaatt aaatcgacta taaaattgtc aatatgttct caattttcca
     3721 gtccaaggcc gccaagacaa gtttgtacaa aaaagcaggc tcgggatccc caaagaagaa
     3781 gcgtaaggtg tccaaggtaa cacttagttt ttgttgctct gtttaaaaat taattttaag
     3841 ggagaggagg acaacatggc ctccctccca gccacccacg agctccacat cttcggctcc
     3901 atcaacggag tcgacttcga catggtcgga caaggaaccg gaaacccaaa cgacggatac
     3961 gaggagctca acctcaagtc caccaaggta agttttcttg agacctttac taactaacta
     4021 ggtctcatag attttcaggg tgacctccaa ttctccccat ggatactcgt cccacacatc
     4081 ggatacggat tccaccaata cctcccatac ccagacggaa tgtccccatt ccaagccgcc
     4141 atggtcgacg ggtccggata ccaagtccac cgtaccatgc aattcgagga cggagcctcc
     4201 ctcaccgtca actaccgtta cacctacgag ggatcgcaca tcaaggtaag ttatgtagag
     4261 acctttacta actaactagg tctcaaacat tttcagggag aggcccaagt caagggaacc
     4321 ggattcccag ccgacggacc agtcatgacc aactccctca ccgccgccga ctggtgccgt
     4381 tccaagaaga cctacccaaa cgacaagacc atcatctcca ccttcaagtg gtcctacacc
     4441 accggaaacg gaaagcgtta ccgttccacc gcccgtacca cctacacctt cgccaagcca
     4501 atggccgcca actacctcaa ggtaagttct ttagagacct ttactaacta actaggtctc
     4561 agtaattttc agaaccaacc aatgtacgtc ttccgtaaga ccgagctcaa gcactccaag
     4621 accgagctca acttcaagga gtggcaaaag gccttcaccg acgtcatggg aatggacgag
     4681 ctctacaagt cccgtcgtcg taaggccaac ccaaccaagc tctccgagaa cgccaagaag
     4741 ctcgccaagg aggtcgagaa cggatccaac ccagctttct tgtacaaagt ggaatagatt
     4801 ctcgacaccc ctaacttcca tcacaagcat tctatttctt tccacaacct tcttaaaccc
     4861 cccgtattcg tttcttccct cacttgtctc tctcacccgc cccgctctcc gacgtcatcc
     4921 accacccatc cattcactaa tttatttgct tttttgattg tttttcttct cggagacgtt
     4981 cgccgtccca agttttatct ctgaaatttg cgtctgagtg aagtgaatgc tgaagtttcg
     5041 tgtagttctc aagatagcgg cccaaaagtg catttgagca caaatatttg tattgtggtt
     5101 ttttgatttc tttaagaagt agccgcaatg ggtttcgaaa tctgttagga tattatgctt
     5161 gtctgagaaa aaaaaacggt gagacgcaga gaaatagtgt ctgcgtctcc acttaatatg
     5221 tgctatttag gctctccgct tcgatatcgc taccgtattc agactgcaaa aaatactcta
     5281 accttttttt tctcagacaa gcataataac aataggtaaa aaaaactgaa cctatttatt
     5341 caaaacttga ttggtaaaac attatataga gaaaccgaaa aatatacaat tgaatcctag
     5401 gttaaattcg ccgccttcac aaatccattc tgctcaactt tattatacat agttgata
//
