LOCUS       sequence-433883-        4934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses rat ⍺SNAP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4934)
  AUTHORS   Leitz J, Wang C, Esquivies L, Peters JJ, Gopal N, Pfuetzner RA, Wang
            AL, Brunger AT
  TITLE     Observing isolated synaptic vesicle association and fusion ex vivo.
  JOURNAL   Nat Protoc. 2024 Jul 2. doi: 10.1038/s41596-024-01014-x.
  PUBMED    38956381
REFERENCE   2  (bases 1 to 4934)
  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: "Nat Protoc.
            2024 Jul 2. doi: 10.1038/s41596-024-01014-x."
COMMENT     Sequence Label: pDC180
FEATURES             Location/Qualifiers
     source          1..4934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     943..960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     944..960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1423..1442
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1423..1441
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1442..1466
                     /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)."
     CDS             1512..1541
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1542..1562
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     terminator      2461..2508
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2731..2750)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(2756..2775)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        2817..2908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2909..2977,2978..3769)
                     /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(3127..3146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3852..4934
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3871..3890)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
      901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
     1021 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
     1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
     1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
     1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
     1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
     1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
     1381 attatcattt agaataaatt ttgtgtcgcc cttccgcgaa attaatacga ctcactatag
     1441 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac ttttaggagg
     1501 taaaacatat gcaccatcac caccaccatc atcatcacca cgagaacctg tattttcaag
     1561 gcatggacac ttcgggtaag caggcagagg cgatggcatt gttggcggag gcagaacgta
     1621 aagttaagaa cagccaaagc tttttcagcg gcctgtttgg tggtagcagc aaaatcgaag
     1681 aggcttgtga gatctacgcg cgtgcagcta atatgttcaa gatggcaaag aattggagcg
     1741 cagcgggtaa tgccttctgt caggccgcac agttgcattt gcaactgcag tctaaacacg
     1801 acgctgccac ctgctttgtg gatgccggca atgctttcaa gaaggcggac ccgcaagagg
     1861 caattaactg tctgatgcgc gcgatcgaga tctataccga catgggtcgt ttcaccattg
     1921 cggctaagca tcatattagc atcgcggaga tttatgaaac cgaactggtc gatgtggaga
     1981 aagcaattgc gcactacgag cagagcgcgg actactacaa gggcgaagag tctaacagca
     2041 gcgcgaacaa atgcctgctg aaagtcgcgg gttatgccgc gcagctggaa caatatcaaa
     2101 aagcgatcga tatctacgag caagttggca cctctgcgat ggacagcccg ctgctgaaat
     2161 actccgccaa agattacttc tttaaagccg cgctgtgcca cttttgcatc gatatgctga
     2221 atgcgaaact ggcagtccag aagtatgaag aattgttccc ggcctttagc gattcccgtg
     2281 agtgcaagct gatgaagaag ctgctggagg cgcacgaaga gcaaaacgtg gatagctata
     2341 ccgaatccgt taaagaatac gatagcatta gccgtctgga ccagtggctg acgacgatgc
     2401 tgctgcgcat taagaaaacg attcagggtg acgaagagga cctgcgctaa ctcgagcccc
     2461 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcc cctgagacgc
     2521 gtcaatcgag ttcgtaccta agggcgacac cccataatta gcccgggcga aaggcccagt
     2581 ctttcgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga
     2641 gtccccacac taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt
     2701 gggaccaccg cgctactgcc gccaggcaaa caaggggtgt tatgagccat attcaggtat
     2761 aaatgggctc gcgataatgt tcagaattgg ttaattggtt gtaacactga cccctatttg
     2821 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2881 gcttcaataa tattgaaaaa ggaagaatat gagtattcaa catttccgtg tcgcccttat
     2941 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3001 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3061 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3121 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3181 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3241 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3301 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3361 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3421 accaaacgac gagcgtgaca ccacgatgcc tgtagcgatg gcaacaacgt tgcgcaaact
     3481 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3541 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3601 taaatccgga gccggtgagc gtggttctcg cggtatcatc gcagcgctgg ggccagatgg
     3661 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3721 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaag cggcgcgcca
     3781 tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     3841 gggtggtgaa tatgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     3901 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     3961 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     4021 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     4081 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     4141 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     4201 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     4261 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     4321 acagtattat tttctcccat gaggacggta cgcgactggg cgtggagcat ctggtcgcat
     4381 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     4441 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     4501 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     4561 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     4621 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     4681 aagatagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     4741 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     4801 agctgttgcc agtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     4861 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4921 aaagcgggca gtga
//
