LOCUS       Exported                7931 bp ds-DNA     circular SYN 24-AUG-2021
DEFINITION  Expresses Protein phosphatase Slingshot homolog 1 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-Ssh1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7931)
  AUTHORS   Aihara S, Fujimoto S, Sakaguchi R, Imai T
  TITLE     BMPR-2 gates activity-dependent stabilization of primary dendrites 
            during mitral cell remodeling.
  JOURNAL   Cell Rep. 2021 Jun 22;35(12):109276. doi: 
            10.1016/j.celrep.2021.109276.
  PUBMED    34161760
REFERENCE   2  (bases 1 to 7931)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 24, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7931
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1733..1742
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3466..3477
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4953..4972)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5018..5073
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5072..5091)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5434..5450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5434..5450)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5447..5469)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5458..5474
                     /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(5482..5512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5527..5548
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5607..5802
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5653..5788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5715..5734
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5808..5942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5845..5864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5899..5918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6010..6027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6181..6769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6261..6280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6940..7800)
                     /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     7563..7582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7801..7905)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 ggccttggtg acccttcagc gctctcccac gcccagcgcc gcgtcttcct ccgcaagcaa
     1801 cagcgagttg gaggctggca gcgatgaaga acggaaattg aacctcagct tgagtgagag
     1861 ctttttcatg gtgaaaggag ctgccctctt cctacagcag ggaaacagcc cacagggcca
     1921 gcggagtctt cagcaccctc acaagcatgc aggtgatctg cctcagcacc tgcaagtgat
     1981 gatcaacctc ctgcgttgtg aagacagaat caagctggcc gtgcgcctag agagtgtctg
     2041 gaccgaccgt gtccgctaca tggtcgtggt atacaccagc gggcgccagg acaccgagga
     2101 gaatattctg ctgggagttg acttttccag taaggagagc aaaagctgca cgatcggaat
     2161 ggttcttcga ctctggagcg acaccaagat tcacctcgat ggggacggcg ggttcagtgt
     2221 gagcacagcg ggcaggatgc acatattcaa gccagtgtct gtccaggcca tgtggtctgc
     2281 cctgcaagtg cttcacaagg cctgcgaagt ggcccggagg cataactact tcccaggagg
     2341 agtggcgctc atctgggcca cctactatga gagctgcatc agctcggagc agagctgcat
     2401 caatgagtgg aatgccatgc aggacctgga gtccacgcgg cccgactccc ccgcgctgtt
     2461 tgtggacaag ccaaccgagg gcgaaagaac tgagcgcctc attaaagcca aactccggag
     2521 catcatgatg agccaggacc ttgaaaatgt gacttctaag gaaatccgaa atgagctgga
     2581 gaagcaaatg aactgcaacc tgaaggagtt caaggaattc atcgataacg agatgctgct
     2641 catcctgggc cagatggaca agccctccct catctttgac catctttatc ttggctccga
     2701 gtggaatgca tccaatctgg aggagctgca gggctcagga gttgactaca ttttaaatgt
     2761 cactagagaa atagacaatt ttttccccgg cttgtttgcg taccataaca tccgcgtgta
     2821 cgatgaggag accacagacc ttcttgccca ctggaatgaa gcttatcatt tcataaacaa
     2881 agcgaaaagg aatcattcca agtgcctggt ccattgcaag atgggcgtca gccgatctgc
     2941 gtccacagtc attgcctacg ccatgaagga gttcggctgg cccctggaga aagcgtataa
     3001 ctacgtgaag cagaaacgta gtatcactcg gcccaacgca ggctttatga ggcagctgtc
     3061 tgagtacgaa ggcatcctgg atgcaagcaa acagcggcat aacaaactat ggcgccagca
     3121 gcccacagat gacaccatcg cagagcccag tgagttcttg ccagagacct tggacggagc
     3181 cctggacgct cagctgccct gtttggatga taccacccac cctgggctcc cgagaagtct
     3241 ggccccagga ggacccgctc tcccctgttg ttttcgaaga ctctcggacc ccctcctcct
     3301 tccccaccat gatgaaacgg gtggcctggt ccacttagag gatctcgaga aggatgctct
     3361 gttagaggag gaggagtctc agccagtgga ggtgcacaag ctggttcagc atccccagga
     3421 aggtgccagg ctgtgtgaaa aggacgtaaa gaggaaactg gagtttggga actccaaacc
     3481 ccgcagtgac tccttgcctc aggtggagga gctggagaag gacggtagcc caagaacggg
     3541 gaggtggagg cgggcctcca cccagctcga tagaagcttg ctcgaccagg aaaacctaaa
     3601 taacaacaac agcaagagga gctgtcccga tgacttggag cgcgatgcca tgtttggaat
     3661 tctcagcaaa gtgaagcctc cctacacatc ctgtgccgac tgcatgtacc ccacagctgg
     3721 tgggactccc gaggcctaca tggagcgaca cgaagacccc agctcctctg ctatctgcac
     3781 ccagccaacc ttcctccccc atgtcacgtc ttccccaatg gcccacgcga gcagcaggtc
     3841 ccgagctccg gagaggccgg cctcgggtcc agccaacacc tctccattcc tactaccagc
     3901 aggctccagg aagccagatg tcagtggctc tggagccggg gctgccccgg aaccaccagc
     3961 aagccttcta gagccttcca gagagacctc aaaagccctt ccaaaatctc tccagttgaa
     4021 gaatcctcac tgtgacaaga acgctgccaa tatggaggtg tcggcgaagg aagaaccgtc
     4081 gcctaagaaa gatcccaagc cggctaagga cctgaggctt ctgttcagta acgaggcgga
     4141 gaagcccacc accaacagct acctgatgca gcatcaggag tccatcatcc aactgcagaa
     4201 ggcaggcctg gtccgaaagc acaccaaaga gctggagagg ttgaagagcc tgccttcaga
     4261 ctcaccagct gcctgcaggg acagcgccac ctgcaggctg gaggccagca tcccggagga
     4321 gggtagccag gagcccgcac acccagccct gtgcagccaa gctgggtcgg aagaacagcc
     4381 tgtaggggga accttgcaga agagccccac gtctaccctc ccccgtttag atcacaccag
     4441 taacttctcg aaggacttcc tgaagaccgt gtgttacacc cccacctcct cctccatcag
     4501 ctccaacctg acccggagct ccagcagcga cagcatccac agcgtccgag ggaagcctgg
     4561 gctggtgaag cagcgggcgc aggagatcga gacgcggctg cgcctggcag gcctcaccgt
     4621 gtcgtccccg ctgaaaaggt cccattccct tgccaagctg ggaagtctca acttctccac
     4681 ggaggacctg tccagcgagg ctgacacatc caccatcgct gactcgcagg acgccaagtg
     4741 tggtctctcc tcttccttct tgcccgaacc ccagtctgcg ccaagggacc ccgctgcaac
     4801 ctctaaatca tcagggaaat ctgccccaga acacttgaaa agcccgtcga gggtaaacaa
     4861 aagctgagcg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     4921 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     4981 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     5041 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     5101 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     5161 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     5221 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     5281 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     5341 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     5401 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5461 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5521 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5581 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     5641 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5701 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5761 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     5821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5941 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6001 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6061 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6121 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6181 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6241 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6301 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6361 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6421 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6481 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6541 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6601 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6661 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6721 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6781 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6841 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6901 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6961 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7021 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7081 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7141 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7201 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7261 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7321 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7381 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7441 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7501 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7561 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7621 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7681 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7741 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7801 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7861 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7921 agtgccacct g
//
