LOCUS       sequence-486788-        5683 bp    DNA     circular SYN 19-SEP-2025
DEFINITION  Expresses rap-1::mCherry in cholinergic motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  AUTHORS   Lamb R, Scales M, Watkins J, Werner M, Cherra SJ
  TITLE     A PDZ-RapGEF promotes synaptic development in C. elegans through a 
            Rap/Rac signaling pathway.
  JOURNAL   Development. 2025 Aug 1:dev.204678. doi: 10.1242/dev.204678.
  PUBMED    40748210
REFERENCE   2  (bases 1 to 5683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development. 2025 Aug 1:dev.204678. doi: 10.1242/dev.204678."
COMMENT     Sequence Label: pBD30
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2692..2716)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2742..2758)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2742..2758)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3495..3515)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3495..3513)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3534..3550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3534..3550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3547..3569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3558..3574
                     /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(3582..3588,3589..3606,3607..3612))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3627..3648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3765..3782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3936..4524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4016..4035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4695..5486,5487..5555))
                     /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     5318..5337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5556..5660)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgc
      661 ttttggtttt cacaattttc tggtttttgg gatttttgcg ggaattttga gaggagaaag
      721 ttgggagaga agatattgga atggtccccg gggagctgta catttataca ccaaggcaat
      781 cttcgtaaaa aataacaata gagtcatccc tcagaacaga aaagaaaccc gcccgggggc
      841 tcaaatacac atgtcatttt tcttttttcg cctgattttt agcctaaatt agtgtttagc
      901 aattttttaa ggtaactcgg tggagtcatt tgaacttttt aaactaaaat ttcagaaaaa
      961 attctttttt tttgtcaaaa aataattttc ccgccggaat ttttcaaaaa aataattttt
     1021 attttatttt tcaagttcct tgtttcctct ttattttttt attttgcccc caaaagtttt
     1081 acactaaaat atttaatttt caggtaattc atcgaaagtc tttctatttt ccgcatctct
     1141 tgttcaaaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat tggccaaagg
     1201 acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag gaggaccctt
     1261 ggagggtacc ggtagaaaaa aagcttgata tcacaagttt gtacaaaaaa gcaggctccg
     1321 aattcgccct tggactagtc atatgggtac catgcgggag tataagattg ttgtgctcgg
     1381 atctggagga gtaggaaaat ccgcactgac ggtacaattc gtgcagggaa ttttcgtaga
     1441 gaagtatgac ccaacaattg aggatagtta cagaaagcaa gtggaagtcg acggccagca
     1501 atgtatgctt gagatactgg atacagcagg aacggagcaa ttcacagcga tgcgtgattt
     1561 gtacatgaag aatggacaag gatttgttct cgtttattca attactgctc aatcgacatt
     1621 caatgatctt atggatcttc gtgatcagat tcttcgtgtc aaagatacgg acgaggttcc
     1681 aatgattttg gtaggaaata agtgtgattt ggaagacgaa cgagttgttg gaaaagatca
     1741 gggacagaat ctcgccaggc agtttggatc agcattcctt gaaacctctg caaaagcaaa
     1801 gattaacgtc agcgaggtgt tctacgacct cgttcgtcaa ataaaccgac gttatcccga
     1861 atctggtcga cgccagggac aatcaaataa gcagtgctgc tcgtgtgtca tcatgggtac
     1921 cagtggttcc ggaggatctg gaggatctgg tatggtctca aagggtgaag aagataacat
     1981 ggcaattatt aaagagttta tgcgtttcaa ggtgcatatg gagggatctg tcaatgggca
     2041 tgagtttgaa attgaaggtg aaggagaagg ccgaccatat gagggaacac aaaccgcaaa
     2101 actaaaggta actaaaggcg gaccattacc attcgcctgg gacatcctct ctccacagtt
     2161 catgtatgga agtaaagctt atgttaaaca tccggcagat ataccagatt atttgaaact
     2221 ttcattcccg gagggtttta agtgggaacg cgtaatgaat tttgaagacg gaggagttgt
     2281 tacagtgacg caagactcaa gcctccaaga tggagaattt atttataaag tcaaacttcg
     2341 aggaacgaat ttcccctcgg atggacctgt tatgcagaag aagactatgg gatgggaagc
     2401 ttcaagtgaa agaatgtacc ctgaagacgg tgctcttaag ggagagatta aacaacgtct
     2461 taaattgaaa gatggaggac attacgatgc tgaggtgaag acaacttaca aagccaaaaa
     2521 accagttcag ctgccaggag cgtacaatgt taatattaaa ctggatatca cctcccacaa
     2581 cgaggattac actatcgttg agcaatatga aagagctgaa gggcggcact cgacaggtgg
     2641 catggatgaa ttgtataagt agcatatggc ggccgcggaa gggcgcattc gacccagctt
     2701 tcttgtacaa agtggtgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     2761 ggccgccacc gcggccgctg tcatcagatc gccatctcgc gcccgtgcct ctgacttcta
     2821 agtccaatta ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt
     2881 tttgttatta tcaaaaaact tctcttaatt tctttgtttt ttagcttctt ttaagtcacc
     2941 tctaacaatg aaattgtgta gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa
     3001 aaattgtgct ccctcccccc attaataata attctatccc aaaatctaca caatgttctg
     3061 tgtacacttc ttatgttttt tacttctgat aaattttttt gaaacatcat agaaaaaacc
     3121 gcacacaaaa taccttatca tatgttacgt ttcagtttat gaccgcaatt tttatttctt
     3181 cgcacgtctg ggcctctcat gacgtcaaat catgctcatc gtgaaaaagt tttggagtat
     3241 ttttggaatt tttcaatcaa gtgaaagttt atgaaattaa ttttcctgct tttgcttttt
     3301 ggggtttccc ctattgtttg tcaagatttc gaggacggcg tttttcttgc taaaatcaca
     3361 agtattgatg agcacgatgc aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg
     3421 aaggtgagta gaagttgata atttgaaagt ggagtagtgt ctatggggtt tttgccgagc
     3481 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     3541 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3601 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3661 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3721 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3781 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3841 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3901 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3961 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4021 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4081 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4141 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4201 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4261 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4321 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4381 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4441 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4501 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4561 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4621 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4681 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4741 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4801 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4861 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4921 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4981 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5041 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5101 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5161 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5221 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5281 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5341 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5401 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5461 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5521 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5581 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5641 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
