LOCUS       sequence-360127-        2319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gibson cloning vector for synthesis of single and double stranded 
            RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2319)
  AUTHORS   Wolff A, Wagner C, Wolf J, Lobo D
  TITLE     In situ probe and inhibitory RNA synthesis using streamlined gene 
            cloning with Gibson assembly.
  JOURNAL   STAR Protoc. 2022 Jun 14;3(3):101458. doi: 
            10.1016/j.xpro.2022.101458. eCollection 2022 Sep 16.
  PUBMED    35733605
REFERENCE   2  (bases 1 to 2319)
  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: "STAR 
            Protoc."; date: "2022-06-14"; volume: "3(3):101458. doi"; pages: " 
            10.1016/j.xpro.2022.101458. eCollection 2022 Sep 16"
COMMENT     Sequence Label: pDL1
FEATURES             Location/Qualifiers
     source          1..2319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(40..87)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     89..105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        126..144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      182..191
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(275..293)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(276..293)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(295..313)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(314..330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(314..330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      332..379
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(386..403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(557..1145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(637..656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1316..2107,2108..2176))
                     /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     1939..1958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2177..2281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc aaaaaacccc tcaagacccg
       61 tttagaggcc ccaaggggtt atgctagggt aaaacgacgg ccagttaata cgactcacta
      121 tagggaatta accctcacta aagggagcgg tgaagagcgg gcccgacgtc gcatgctccc
      181 ggccgccatg gccgcgggat atcactagtg cggccgcctg caggtcgacc atatgggaga
      241 gctcccaacg cgttggatgc atgctcttcg cgcttctata gtgtcaccta aatccctata
      301 gtgagtcgta ttagtcatag ctgtttcctg tctagcataa ccccttgggg cctctaaacg
      361 ggtcttgagg ggttttttgc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      421 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      481 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      541 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      601 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      661 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      721 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      781 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      841 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      901 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      961 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1021 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1081 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1141 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1201 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1261 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1321 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1381 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1441 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1501 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1561 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1621 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1681 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1741 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1801 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1861 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1921 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1981 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2041 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2101 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2161 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2221 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2281 gcacatttcc ccgaaaagtg ccacctgatg cggtgtgaa
//
