LOCUS       sequence-483115-        7802 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Expresses tiam-1::mGFP in cholinergic motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7802)
  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 7802)
  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: pBD34
FEATURES             Location/Qualifiers
     source          1..7802
                     /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"
     CDS             join(4062..4064,4065..4067,4068..4781)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4107..4128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4368..4387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4715..4736
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4811..4835)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4861..4877)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4861..4877)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5614..5634)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5614..5632)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5653..5669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5653..5669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5666..5688)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5677..5693
                     /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(5701..5707,5708..5725,5726..5731))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5746..5767
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5884..5901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6055..6643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6135..6154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6814..7605,7606..7674))
                     /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     7437..7456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7675..7779)
                     /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 gggctcacgc ctctcatgtt
     1381 cttgctcaca agccgaatgc tggacgaatg atgaatctac attgagaatt gacgcggagt
     1441 tgtttgagtt atcacaggac ggacaaaagt gggataagct tgacggaagg ctggccaacg
     1501 tacgtgtatt caacgcattc gacgactctt ccccacgtct tcttgccaca tcatcatccg
     1561 gtcaagttct tctggacact ctaattccaa tgggagaaaa ggttcataaa gtcagcgact
     1621 tttttgtgta tttgaagacg gaaggaagaa caattggatt caatacgtta agctcgaggg
     1681 acacttcctt actcatttct caggcgacaa atacaactgc cttcaatatg tttcaacatt
     1741 cattctcagt ttccgtatcc agaattgcta catttgaagg tgattctatt caaccttgtg
     1801 atgagcagcc agaaacgtca aatgtctcac taaatccgtt aaatatggtt ttcaactaca
     1861 aaggatcacc tgttgagatt gatttgctgg aatgtttcgc gtgctcatca gaagctgaaa
     1921 actgtgtcga catagcacat gccaacaaca tatttcgtgt aattgtgtca gcacatgctc
     1981 atctttttct tattcaagct ctgaatgtat catcacttct tcttgctcga accacctcga
     2041 atctagttgc catcaattat ttggctcggc aaaatgataa aattcaacaa aaattgattg
     2101 ctctgttgga aacatcagaa tcaagtgaga atgcgtcgaa ggaagatatt cgacaaatgc
     2161 ttcttctgaa gaatgtagta gtgacacaga gattgaactc acttcaacag agagcactac
     2221 cgggaattga gcttatccaa cacgatatct cacctcaaat tgatcagaag ctactcgaaa
     2281 agtgtcgact tccatcactg tttttatatc attttaatat gaattcgttt cgaatatggc
     2341 gaaaaccaat ggaaacacgt gacatgaaca caatgggatc atcaatggca tccaccagtc
     2401 aataccatca gattccgatt caaatccaga atcagaatcc gttaccgtct caacagattc
     2461 agccacaaca acaacaacaa taccatccac cagatcatgt gacaactcat aatggtacaa
     2521 ctacgtttca tactacgaat cccacaaaac caagactttg tgaaggatcg ttggctcctg
     2581 gatgctcatt ggccgaggca tctgaacaga gaaaacgaga gatggcggag cagaaaaaag
     2641 agttggcagg cccatcttcc gcagaggatt acttcgtcag aaagacaaat ggacgtcttg
     2701 gactcactat atacgcgcat aacgatgatg gggtcatcag ggctgaggtt cgtggagtga
     2761 cgtcatttgc tccgagatgt gctcaagttg gagattcggt tgtagcggta gacagtgaac
     2821 tcatatcatc agttagaaac gcttcggatg ttgagaaact tttgagaatc ggaaaagtaa
     2881 tacatttacg aagaaagact cctctaactc caagagcacc gtctcaactc gacgcaacta
     2941 atgtcaataa gaaaacgggc tgtgcaaagt tggcgatggc tctgcaagaa ttgttggtca
     3001 ctgagaagaa atatgtcagc gatcttcgag agatgaatga agtttttctg tgtatgagac
     3061 aagtccgaga tattatgaat gcagcaatac ggttgtataa aatgcaaaca tcgtttgtgg
     3121 acagtattga ggaagccatt ggagacatga gtagacagga tatttctgtt gctcagatta
     3181 gggattctgt aatgcgagtg tgtgcagtat tcatcaacaa atgttctgat ttcaaaatct
     3241 atgcagaata tgcagccggt taccatctcc tccaacatga gatcaaaaac aaaaaagagc
     3301 ttctatcaaa attggaagct gtaaacagta caagagagca acattgttct tgggaatctc
     3361 ggatgattaa accagtccag agaattgtac aatatccact tttactgaaa aacattgccg
     3421 atgcacttcc gaaggatgca cgagagaggg ttcaagtgga ggcagcattg caaaaaatgc
     3481 agacgtctgc agaatatgtg aatgaaatgc aaaggcttca tgaagattat gcacaatata
     3541 tggagactgt taggaaagct aatgagccaa tgttggctga aaagagtctt cgcctcgaca
     3601 ctcgagagct tctagttttt gcacacataa aatggagaga tgctccagcg gagcactacg
     3661 ttgtgttcgt gtttcactct ttaatactgc tccttccatc gtatgctagg aaagagatta
     3721 agatgaaatg gactcgtgtc ctcccaataa atgaagttga tatcaacgaa atgccaaatg
     3781 atgcattaaa tctaaaactt tatcatgctg cttttgaagg accaaatgga caattaagtc
     3841 attccaatcc aaatacagtc tacaatattg aatgttgtca atctcaactg aaacagcatc
     3901 ttatcaaaaa tatcaaaaaa gctcgaactt tattctctcg agaatcacat cgtccattaa
     3961 gtggatctag tcaatcggat ggtggttatg tgtcagaagt ctctaaggaa catcgaaatt
     4021 caaaaggtac cagtggttcc ggaggatctg gaggatctgg tatggtgagc aagggcgagg
     4081 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4141 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4201 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     4261 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4321 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4381 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4441 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4501 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4561 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4621 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     4681 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4741 ccgccgggat cactctcggc atggacgagc tgtacaagta gcatatggcg gccgcggaag
     4801 ggcgaattcg acccagcttt cttgtacaaa gtggtgatat cgaattcctg cagcccgggg
     4861 gatccactag ttctagagcg gccgccaccg cggccgctgt catcagatcg ccatctcgcg
     4921 cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc
     4981 tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt
     5041 tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt
     5101 cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca
     5161 aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg
     5221 aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg
     5281 accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg
     5341 tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat
     5401 tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt
     5461 ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt
     5521 tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc
     5581 tatggggttt ttgccgagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     5641 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5701 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5761 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5821 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5881 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5941 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6001 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6061 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6121 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6181 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6241 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6301 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6361 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6421 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6481 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6541 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6601 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6661 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6721 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6781 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6841 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6901 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6961 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7021 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7081 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7141 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7201 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7261 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7321 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7381 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7441 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7501 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7561 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7621 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7681 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7741 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7801 ac
//
