LOCUS       pHnS_2comp_KI;Nlgn2_#2        6469 bp    DNA     circular UNA 11-JUL-2025
DEFINITION  .
ACCESSION   urn.local...1b-jebled8
VERSION     urn.local...1b-jebled8
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..142
                     /standard_name="ITR"
     source          181..1067
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    181..200
                     /standard_name="Primer site #2"
     regulatory      201..513
                     /note="Geneious type: promoter"
                     /note="RNA polymerase III promoter for mouse U6 snRNA (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/3366121""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/3366121"">Das
                     et al., 1988</a>)"
                     /standard_name="mU6 promoter"
     primer_bind     432..452
                     /standard_name="mU6-F"
     primer_bind     complement(493..514)
                     /standard_name="SEQ_mU6_last_R"
     misc_feature    515..534
                     /locus_tag="DRSR2(1)"
                     /standard_name="DRSR2"
     misc_feature    535..630
                     /standard_name="Scaffold cr2"
     misc_feature    597..608
                     /standard_name="T-lock"
     misc_feature    631..637
                     /standard_name="polyT"
     misc_feature    638..656
                     /standard_name="Primer site #3"
     misc_feature    673..913
                     /standard_name="hU6"
     primer_bind     673..693
                     /standard_name="hU6-F"
     primer_bind     844..863
                     /standard_name="LKO.1 5'"
     misc_feature    923..942
                     /standard_name="Nlgn2#2"
     misc_feature    943..959
                     /standard_name="Scaffold cr3"
     misc_feature    964..1038
                     /standard_name="Scaffold cr3"
     misc_feature    1005..1016
                     /standard_name="T-lock"
     misc_feature    1039..1045
                     /standard_name="polyT"
     misc_feature    1046..1067
                     /standard_name="Primer site #4"
     regulatory      1099..1546
                     /note="Geneious type: promoter"
                     /note="human synapsin I promoter; confers neuron-specific
                     expression (<a
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/12595892""
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/12595892"">Kügl
                     er et al., 2003</a>)"
                     /standard_name="hSyn promoter"
     primer_bind     complement(1100..1123)
                     /standard_name="SEQ_hSyn_0R"
     CDS             1576..2644
                     /codon_start=1
                     /locus_tag="smFP HA"
                     /transl_table=1
                     /translation="MYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGGVSKGEELFTGVV
                     PILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLGGGVQC
                     FSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGI
                     DFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEGGYPYDVPDYAGGY
                     PYDVPDYAGGGGYPYDVPDYAGGYPYDVPDYAGGDGSVQLADHYQQNTPIGDGPVLLP
                     DNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGYPYDVPDYAGYPYD
                     VPDYAGYPYDVPDYA*TGARGT"
                     /standard_name="smFP HA"
     CDS             1579..1605
                     /codon_start=1
                     /locus_tag="HA(5)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             1609..1635
                     /codon_start=1
                     /locus_tag="HA(6)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             1639..1665
                     /codon_start=1
                     /locus_tag="HA(7)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2194..2220
                     /codon_start=1
                     /locus_tag="HA(8)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2227..2253
                     /codon_start=1
                     /locus_tag="HA"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2266..2292
                     /codon_start=1
                     /locus_tag="HA(1)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2299..2325
                     /codon_start=1
                     /locus_tag="HA(2)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2536..2562
                     /codon_start=1
                     /locus_tag="HA(3)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2566..2592
                     /codon_start=1
                     /locus_tag="HA(4)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     CDS             2596..2622
                     /codon_start=1
                     /locus_tag="HA(9)"
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /standard_name="HA"
     misc_feature    2657..2671
                     /locus_tag="X"
                     /standard_name="X"
     misc_feature    2686..3274
                     /locus_tag="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /standard_name="WPRE"
     primer_bind     complement(2739..2759)
                     /locus_tag="WPRE-R"
                     /note="WPRE, reverse primer"
                     /standard_name="WPRE-R"
     CDS             complement(3157..3168)
                     /codon_start=1
                     /locus_tag="Factor Xa site"
                     /product="Factor Xa recognition and cleavage site"
                     /transl_table=1
                     /translation="IEGR"
                     /standard_name="Factor Xa site"
     primer_bind     complement(3277..3294)
                     /locus_tag="BGH-rev"
                     /note="Bovine growth hormone terminator, reverse primer.
                     Also called BGH reverse"
                     /standard_name="BGH-rev"
     misc_feature    3283..3490
                     /note="Geneious type: polyA_signal"
                     /locus_tag="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
                     /standard_name="bGH poly(A) signal"
     misc_feature    3533..3674
                     /standard_name="ITR"
     primer_bind     complement(3699..3715)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /standard_name="M13 rev"
     protein_bind    3723..3739
                     /bound_moiety="<i>lac</i> repressor encoded by
                     <i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli</i>. This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /standard_name="lac operator"
     regulatory      join(complement(3772..3777),complement(3754..3771),
                     complement(3747..3753))
                     /note="Geneious type: promoter"
                     /note="promoter for the <i>E. coli lac</i> operon"
                     /standard_name="lac promoter"
     protein_bind    3792..3813
                     /bound_moiety="<i>E. coli</i> catabolite activator
                     protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /standard_name="CAP binding site"
     rep_origin      complement(4101..4689)
     CDS             join(complement(5652..5720),complement(4860..5651))
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /standard_name="AmpR"
     regulatory      complement(5721..5825)
                     /note="Geneious type: promoter"
                     /gene="<i>bla</i>"
                     /standard_name="AmpR promoter"
     rep_origin      complement(5851..6306)
ORIGIN      
        1 aggttaatta aggctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg
       61 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
      121 caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac
      181 ccagtctgac tatcacgatg gatccgacgc gccatctcta ggcccgcgcc ggccccctcg
      241 cacagacttg tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt
      301 tcctagtaac tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata
      361 ctagctacat tttacatgat aggcttggat ttctataaga gatacaaata ctaaattatt
      421 attttaaaaa acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga
      481 gactataaat atcccttgga gaaaagcctt gttggcgatc gtaatcaccc gagtgtttga
      541 gagctatgca gaaagctgca tagcaagttc aaataaggct agtccgtaca caacttgggc
      601 ttcggcctaa gtggcagccg agtcggctgc tttttttgct cgaatctaca ctcagcgcgg
      661 ccgctcacat gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag
      721 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
      781 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
      841 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
      901 ttgtggaaag gacgaaacac cgccactcca ccactcgggt atgtttcaga gctatgcaca
      961 agagtgcata gcaagttgaa ataaggctag tccgtttaca acttggagct tgctccaagt
     1021 ggcacccgag tcgggtgctt tttttctcga gtactaggat ccattaggcg gtctagactg
     1081 cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
     1141 tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat
     1201 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
     1261 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
     1321 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     1381 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     1441 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     1501 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga gaaggtaccg
     1561 gatccggccg ccaccatgta cccttatgat gtgcccgatt atgctggcta cccttatgat
     1621 gttcctgatt acgccggata tccgtatgac gtgccagact acgcgggagg tgtgagcaag
     1681 ggagaggagt tgtttaccgg cgtggtcccc atactggtgg agttggatgg cgacgttaat
     1741 ggacataaat tctcggtccg cggcgaggga gagggagacg ccaccaatgg caagctgacg
     1801 cttaagttca tttgcacgac tggaaaattg cctgtcccct ggccgaccct ggtcaccaca
     1861 ctgggcggag gagtgcagtg cttctcccgt tacccagacc acatgaagca gcacgatttc
     1921 ttcaagagcg caatgccgga gggatacgtg caagaacgga ctatttcctt caaggatgac
     1981 ggaacctata agacccgtgc ggaggtcaaa ttcgagggtg acaccctggt gaaccgaatt
     2041 gaactcaaag gaatcgattt caaggaggat ggaaatattc tgggtcacaa gctggaatac
     2101 aacttcaaca gccataatgt gtacattacg gctgataagc agaagaacgg cattaaggcc
     2161 aatttcaaga tccgccacaa cgttgagggt ggatatccct acgacgtgcc cgattatgcg
     2221 ggcggatatc cgtatgatgt tccagattat gctggtggtg gaggctatcc ctatgatgtc
     2281 cccgactacg ccggaggtta cccatacgac gtgcccgact acgctggagg cgatggcagc
     2341 gtgcagctgg cagatcatta tcaacagaat acccccatag gtgatggccc cgttctgctt
     2401 ccagataatc actacctttc cacccagagc gtgctttcga aagacccgaa cgaaaagcgt
     2461 gatcacatgg tcctgctgga gtttgtgacc gcggctggaa tcaccctggg tatggacgaa
     2521 ctctacaagg gtggctaccc ctacgatgtg ccggattacg ctggatatcc gtatgacgta
     2581 ccggactatg ccggttaccc gtacgatgtc ccggactacg cttaaaccgg tgctcgaggt
     2641 accgaattcg atatcatagc tagctagcta gggatcggtg tcgacaatca acctctggat
     2701 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2761 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2821 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2881 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2941 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3001 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3061 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     3121 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3181 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtctacgcct tcgccctcag
     3241 acgagtcgga tctccctttg ggccgcctcc ccgcctcctc gactgtgcct tctagttgcc
     3301 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3361 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3421 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc
     3481 atgctgggga gcgggtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     3541 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3601 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3661 gcctaattaa ggccttaatt aaatctggcg taatcatggt catagctgtt tcctgtgtga
     3721 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3781 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3841 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3901 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3961 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4021 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4081 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4141 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4201 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4261 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4321 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4381 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4441 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4501 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4561 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4621 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4681 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4741 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4801 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4861 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4921 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4981 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5041 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5101 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5161 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5221 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5281 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5341 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5401 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5461 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5521 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5581 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5641 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5701 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5761 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5821 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     5881 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5941 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     6001 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     6061 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     6121 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     6181 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     6241 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     6301 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6361 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     6421 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
