LOCUS       sequence-483113-        5690 bp    DNA     circular SYN 19-AUG-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 5690)
  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 5690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 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..5690
                     /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    1300..1324
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2699..2723)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2749..2765)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2749..2765)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3502..3522)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3502..3520)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3541..3557)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3541..3557)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3554..3576)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3565..3581
                     /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(3589..3595,3596..3613,3614..3619))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3634..3655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3772..3789)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3943..4531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4023..4042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4702..5493,5494..5562))
                     /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     5325..5344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5563..5667)
                     /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 tctagcgcca
      781 aggcaatctt cgtaaaaaat aacaatagag tcatccctca gaacagaaaa gaaacccgcc
      841 cgggggctca aatacacatg tcatttttct tttttcgcct gatttttagc ctaaattagt
      901 gtttagcaat tttttaaggt aactcggtgg agtcatttga actttttaaa ctaaaatttc
      961 agaaaaaatt cttttttttt gtcaaaaaat aattttcccg ccggaatttt tcaaaaaaat
     1021 aatttttatt ttatttttca agttccttgt ttcctcttta tttttttatt ttgcccccaa
     1081 aagttttaca ctaaaatatt taattttcag gtaattcatc gaaagtcttt ctattttccg
     1141 catctcttgt tcaaaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggattgg
     1201 ccaaaggacc caaaggtatg tttcgaatga tactaacata acatagaaca ttttcaggag
     1261 gacccttgga gggtaccggt agaaaaaaag cttgatatca caagtttgta caaaaaagca
     1321 ggctccgaat tcgcccttgg actagtcata tgggtaccat gcgggagtat aagattgttg
     1381 tgctcggatc tggaggagta ggaaaatccg cactgacggt acaattcgtg cagggaattt
     1441 tcgtagagaa gtatgaccca acaattgagg atagttacag aaagcaagtg gaagtcgacg
     1501 gccagcaatg tatgcttgag atactggata cagcaggaac ggagcaattc acagcgatgc
     1561 gtgatttgta catgaagaat ggacaaggat ttgttctcgt ttattcaatt actgctcaat
     1621 cgacattcaa tgatcttatg gatcttcgtg atcagattct tcgtgtcaaa gatacggacg
     1681 aggttccaat gattttggta ggaaataagt gtgatttgga agacgaacga gttgttggaa
     1741 aagatcaggg acagaatctc gccaggcagt ttggatcagc attccttgaa acctctgcaa
     1801 aagcaaagat taacgtcagc gaggtgttct acgacctcgt tcgtcaaata aaccgacgtt
     1861 atcccgaatc tggtcgacgc cagggacaat caaataagca gtgctgctcg tgtgtcatca
     1921 tgggtaccag tggttccgga ggatctggag gatctggtat ggtctcaaag ggtgaagaag
     1981 ataacatggc aattattaaa gagtttatgc gtttcaaggt gcatatggag ggatctgtca
     2041 atgggcatga gtttgaaatt gaaggtgaag gagaaggccg accatatgag ggaacacaaa
     2101 ccgcaaaact aaaggtaact aaaggcggac cattaccatt cgcctgggac atcctctctc
     2161 cacagttcat gtatggaagt aaagcttatg ttaaacatcc ggcagatata ccagattatt
     2221 tgaaactttc attcccggag ggttttaagt gggaacgcgt aatgaatttt gaagacggag
     2281 gagttgttac agtgacgcaa gactcaagcc tccaagatgg agaatttatt tataaagtca
     2341 aacttcgagg aacgaatttc ccctcggatg gacctgttat gcagaagaag actatgggat
     2401 gggaagcttc aagtgaaaga atgtaccctg aagacggtgc tcttaaggga gagattaaac
     2461 aacgtcttaa attgaaagat ggaggacatt acgatgctga ggtgaagaca acttacaaag
     2521 ccaaaaaacc agttcagctg ccaggagcgt acaatgttaa tattaaactg gatatcacct
     2581 cccacaacga ggattacact atcgttgagc aatatgaaag agctgaaggg cggcactcga
     2641 caggtggcat ggatgaattg tataagtagc atatggcggc cgcggaaggg cgaattcgac
     2701 ccagctttct tgtacaaagt ggtgatatcg aattcctgca gcccggggga tccactagtt
     2761 ctagagcggc cgccaccgcg gccgctgtca tcagatcgcc atctcgcgcc cgtgcctctg
     2821 acttctaagt ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc
     2881 ccctattttt gttattatca aaaaacttct cttaatttct ttgtttttta gcttctttta
     2941 agtcacctct aacaatgaaa ttgtgtagat tcaaaaatag aattaattcg taataaaaag
     3001 tcgaaaaaaa ttgtgctccc tccccccatt aataataatt ctatcccaaa atctacacaa
     3061 tgttctgtgt acacttctta tgttttttac ttctgataaa tttttttgaa acatcataga
     3121 aaaaaccgca cacaaaatac cttatcatat gttacgtttc agtttatgac cgcaattttt
     3181 atttcttcgc acgtctgggc ctctcatgac gtcaaatcat gctcatcgtg aaaaagtttt
     3241 ggagtatttt tggaattttt caatcaagtg aaagtttatg aaattaattt tcctgctttt
     3301 gctttttggg gtttccccta ttgtttgtca agatttcgag gacggcgttt ttcttgctaa
     3361 aatcacaagt attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc
     3421 tcagtggaag gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt
     3481 gccgagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     3541 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     3601 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     3661 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     3721 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3781 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3841 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3901 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3961 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4021 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4081 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4141 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4201 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4261 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4321 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4381 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4441 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4501 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4561 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4621 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4681 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4741 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4801 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4861 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4921 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4981 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5041 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5101 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5161 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5221 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5281 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5341 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5401 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5461 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5521 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5581 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5641 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
