LOCUS       sequence-490244-        3254 bp    DNA     circular SYN 22-OCT-2025
DEFINITION  For making other mouse ADF silencing vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3254)
  TITLE     Bamburg lab plasmids 2025
REFERENCE   2  (bases 1 to 3254)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: ADF shRNA mouse
FEATURES             Location/Qualifiers
     source          1..3254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..96
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     88..105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     89..105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     115..134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        115..133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     166..182
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     166..182
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        232..446
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     367..386
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(555..575)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(555..573)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(594..610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    618..634
                     /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(642..648,649..666,667..672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    687..708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(825..842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(996..1584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1076..1095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1755..2546,2547..2615))
                     /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     2378..2397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2616..2720)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2747..3202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3044..3065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
       61 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
      121 gactcactat agggcgaatt ggagctccac cgcggtggcg gccgctctag aactagtgga
      181 tccactagta acggccgcca gtgtgctgga attcgccctt ccatggaatt cgaacgctga
      241 cgtcatcaac ccgctccaag gaatcgcggg cccagtgtca ctaggcggga acacccagcg
      301 cgcgtgcgcc ctggcaggaa gatggctgtg agggacaggg gagtggcgcc ctgcaatatt
      361 tgcatgtcgc tatgtgttct gggaaatcac cataaacgtg aaatgtcttt ggatttggga
      421 atcttataag ttctgtatga gaccacagat ctaagtgatt gcaatccgtg tatttctcga
      481 gaatacacgg attgcaatca ctttttttgg aaaagcttgg tcatagctgt ttccggggta
      541 cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
      601 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
      661 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
      721 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
      781 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
      841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1381 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1681 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1741 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1801 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     1861 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     1921 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     1981 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2041 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2101 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2161 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2221 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2281 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2341 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2401 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2461 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2521 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2581 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2701 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     2761 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2821 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2881 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2941 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3001 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3061 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3121 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3181 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     3241 gatcggtgcg ggcc
//
