LOCUS       Exported                5515 bp ds-DNA     circular SYN 17-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5515)
  AUTHORS   Vivek K Sahoo
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5515
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..183
                     /gene="miscellaneous"
                     /label=ITR
                     /note="ITR"
                     /note="/SECDrawas=Region"
     CDS             1
                     /label=accession:
                     /note="accession: "
                     /note="/SECDrawas=Gene"
                     /note="/organism="
     promoter        237..708
                     /gene="promoter"
                     /note="Synapsin-1"
                     /note="/SECDrawas=Region"
     promoter        258..705
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     primer_bind     644..664
                     /label=O3360
                     /note="Name of container: Oligo Box 23O3360-0 (Currently 
                     set Isolate Being Used)H : 6"
     primer_bind     720..751
                     /label=BamHI-EGFP_s
     regulatory      732..741
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             738..1457
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(783..804)
                     /label=O1098
     primer_bind     1045..1064
                     /label=O3312
     primer_bind     complement(1152..1171)
                     /label=O3313
     primer_bind     1287..1304
                     /label=O3339
     primer_bind     1320..1337
                     /label=O3301
     primer_bind     complement(1335..1355)
                     /label=O3340
     primer_bind     complement(1395..1413)
                     /label=O3302
     primer_bind     1408..1472
                     /label=O3553_Dec_UMI27_read1_fwd
     primer_bind     1413..1472
                     /label=O3551_Dec_UMI23_read1_fwd
     primer_bind     1415..1472
                     /label=O3552_Dec_UMI21_read1_fwd
     primer_bind     1421..1472
                     /label=O3303_Dec_UMI25_read1_fwd
     primer_bind     complement(1433..1537)
                     /label=shRNA_scRNA_Perturb2_as
     primer_bind     1462..1483
                     /label=Partial Read2
                     /note="For PCR1(Read amplification PCR)"
     primer_bind     1463..1483
                     /label=O3456_Dec_read2_frame2_rev(-T)
     primer_bind     1464..1483
                     /label=O3458_Dec_read2_frame4_rev(-T)
     primer_bind     1464..1483
                     /label=O3457_Dec_read2_frame3_rev(-T)
     protein_bind    complement(1482..1491)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1482..1483
                     /gene="3' cloning site"
                     /label=attB2 OpenFreezer Core Recombination Site
                     /note=" attB2 OpenFreezer Core Recombination Site"
                     /note="/SECDrawas=Region"
     gap             1484..1503
                     /estimated_length=20
     primer_bind     complement(1504..1553)
                     /label=O3553_Dec_UMI27_read1_fwd
     primer_bind     complement(1504..1551)
                     /label=O3303_Dec_UMI25_read1_fwd
     primer_bind     complement(1504..1550)
                     /label=O3551_Dec_UMI23_read1_fwd
     primer_bind     complement(1504..1548)
                     /label=O3552_Dec_UMI21_read1_fwd
     promoter        complement(1504..1522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1510..1526)
                     /label=O1490
     polyA_signal    1549..1670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(1857)
                     /gene="miscellaneous"
                     /label=shRNA stuffer
                     /note="shRNA stuffer"
                     /note="/SECDrawas=Region"
     promoter        complement(1862..2123)
                     /gene="promoter"
                     /note="hU6"
                     /note="/SECDrawas=Region"
     promoter        complement(1867..2107)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     1908..1935
                     /label=O1459
     primer_bind     complement(2037..2063)
                     /label=O1460
     misc_feature    complement(2210..2392)
                     /gene="miscellaneous"
                     /label=ITR
                     /note="ITR"
                     /note="/SECDrawas=Region"
     promoter        complement(2393..2722)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2407..2542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(2968..3556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3727..4587)
                     /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"
     promoter        complement(4588..4692)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5480..5498
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc tgagctcgct agcctcgagg atatcaagat ctggcctcgg cggccactgc
      241 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
      301 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
      361 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
      421 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
      481 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
      541 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
      601 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
      661 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgaa gctgctagca
      721 aggatccacc cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      781 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      841 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      901 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      961 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1021 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1081 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1141 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1201 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1261 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1321 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1381 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1441 acgagctgta caagtaaaag cttcactaaa gggtaggtga cacnnnnnnn nnnnnnnnnn
     1501 nnncctatag tgagtcgtat tagtagcggc cgcttcgagc agacatgata agatacattg
     1561 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1621 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1681 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     1741 aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagtaaaaaa gaattaaaag
     1801 ctgacgtatc cttaatgtgt gctatgaata ttaacacaca ttaaggatac gtcagcccgg
     1861 tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg
     1921 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
     1981 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
     2041 caattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
     2101 ggccctcggt gaagggggcg gccatcgatg aagacaatag caggcatgct ggggatgcgg
     2161 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgactaga gcatggctac
     2221 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     2281 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2341 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagagc tttttgcaaa
     2401 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
     2461 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     2521 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     2581 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
     2641 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     2701 ttccacaccc taactgacac acattccaca gctgcattaa tgaatcggcc aacgcgcggg
     2761 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2821 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2881 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2941 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3001 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3061 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3121 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3181 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3241 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3301 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3361 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3421 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3481 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3541 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3601 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3661 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3721 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3781 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3841 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3901 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3961 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4021 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4081 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4141 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4201 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4261 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4321 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4381 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4441 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4501 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4561 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4621 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4681 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     4741 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4801 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4861 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4921 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc attcgacgct
     4981 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     5041 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     5101 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     5161 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     5221 tgccggccac gatgcgtccg gcgtagagga tctggctagc gatgaccctg ctgattggtt
     5281 cgctgaccat ttccgggtgc gggacggcgt taccagaaac tcagaaggtt cgtccaacca
     5341 aaccgactct gacggcagtt tacgagagag atgatagggt ctgcttcagt aagccagatg
     5401 ctacacaatt aggcttgtac atattgtcgt tagaacgcgg ctacaattaa tacataacct
     5461 tatgtatcat acacatacga tttaggtgac actatagaat acacggaatt aattc
//
