LOCUS       Exported                3505 bp ds-DNA     circular SYN 27-OCT-2021
DEFINITION  Multisite gateway middle entry clone containing zebrafish 
            Cavin4b_PRDmt with no stop codon. Parton lab clone JLY..
ACCESSION   .
VERSION     .
KEYWORDS    pME-Cavin4b_PRDmt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3505)
  TITLE     Harriet's Cavin4 paper
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3505
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      669..678
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(1630..1729)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1746..1765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1747..1765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1770..1786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1770..1786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1888..1907)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1899..2708
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1966..1985)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2855..3443
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3344..3363
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc caccatggct gataaactgg gactggcagg tgtggcggac gagcccagcg
      721 ccatgaacat cttctctctg ctggagaggg tgtccgggat catagacaat gttcaggcct
      781 gccagcagcg gatggaggag aggcagctgg agctggagaa caccgtcaag accatccaag
      841 cagatgtagt caagctgacg aaggagcaca cggcgaccag tggcaccgtg gagaaacttc
      901 tggagaaaac acgcaaggtc agctgccaca tcaaggacgt ccgcgtgcgg gtggagaagc
      961 agaacgttcg agtcaagaag gtggaggaaa cccaagtcga gctgctgaat aagaacaagt
     1021 tccgtgtcgt catctaccag ggagataatg aggtcccagc tgtggctggt cctaaagcct
     1081 ctccgaaggg agcaatgggt ggagacgctg ctgcagaccc agatgcacca gaggctccgc
     1141 tccctgactc ggatgaggaa tacatggttg ttgaagaagc cgattcttcc actgctgcca
     1201 aactaaagaa gaccggcctg aagcgcatcg aaagcctgaa gcagaccttc tcacgcgaga
     1261 acatgactaa gaccaaagaa aacctgggca ccaaggtcaa caagctgggc gagcgcatag
     1321 taacagccga acggcgtgag aaaatccgcc agtcaggaga acgactgaaa cagtctggcg
     1381 agcgcttcaa ggaaaccatc acaaaaaccg taccagcaaa gctgaatctg aagaaggaaa
     1441 ggacggtggc tgagggtcag gagggggcag aagggaccac agaaggaaca gcagccgtca
     1501 caccaccgaa gggtcgtaaa cccagtcctg aggtggccta cacagagcca acgcagaaac
     1561 aagaaggtcc agatgagggc aagggtgagg aatcagaagt accgatgtat gatatgaagc
     1621 agctttctga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1681 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1741 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1801 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1861 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     1921 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1981 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     2041 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     2101 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     2161 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     2221 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     2281 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     2341 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     2401 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     2461 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     2521 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     2581 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     2641 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     2701 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2761 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3181 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3241 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3301 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3361 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3421 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3481 tttgctggcc ttttgctcac atgtt
//
