LOCUS       Exported                5679 bp ds-DNA     circular SYN 24-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  AUTHORS   lab of neurogenetics
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 4, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5679
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      670..679
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             679..708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             724..753
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    772..796
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    814..838
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     852..871
                     /label=Fragment.FOR
     primer_bind     853..871
                     /label=RILPL1_F
     primer_bind     853..871
                     /label=Insert.FOR
     primer_bind     complement(2040..2064)
                     /label=Fragment.REV
     primer_bind     complement(2040..2064)
                     /label=Insert.REV
     primer_bind     complement(2040..2061)
                     /label=RILPL1_R
     protein_bind    complement(2078..2102)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2181..2199)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2473..2594
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2601..3056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3083..3187
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3189..3546
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3397..3532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3581..4375
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4607..4654
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4983..5571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccatgga gcagaaactc atctctgaag aggatctgag cccgggcgga
      721 tccgagcaga aactcatctc tgaagaggat ctgctgcagg aattcgatat cacaagtttg
      781 tacaaaaaag caggctccga attcgccctt atcacaagtt tgtacaaaaa agcaggctcc
      841 gaattcgccc ttatggagga ggagcggggg tcggcgctgg cggccgagtc ggcgctggag
      901 aagaacgtgg ccgagctgac cgtcatggac gtgtacgaca tcgcgtcgct tgtgggccac
      961 gagttcgagc gggtcattga ccagcacggc tgcgaggcca tcgcgcgcct catgcccaag
     1021 gtcgtgcgcg tcctggagat cctggaggtg ctggtcagcc gccaccacgt cgcgcccgag
     1081 ctggacgagc tgcgcctgga gctggaccgc ctgcgcctgg agaggatgga ccgcatcgag
     1141 aaggagcgca agcaccagaa ggagctggag ctggtggagg atgtgtggcg aggggaggcg
     1201 caggacctcc tctcccagat cgcccagctg caggaggaga acaagcagct catgaccaac
     1261 ctctcccaca aggatgtcaa cttctcagag gaggagttcc agaagcatga aggcatgtca
     1321 gagcgggagc gacaggtgat gaagaagctg aaggaggtgg tggacaaaca acgcgacgag
     1381 atccgcgcca aggacaggga gctgggcctg aaaaatgagg acgttgaggc tttacagcag
     1441 cagcagacac ggctgatgaa gatcaaccat gaccttcggc accgggtcac ggtggtggag
     1501 gcccagggga aagccctgat cgaacagaag gtggagctgg aggcagacct gcagaccaag
     1561 gagcaggaga tgggcagcct gcgagcagag ctggggaagt tgcgagagag gctgcagggg
     1621 gagcacagcc agaatgggga ggaggagcct gagacggagc cggtgggaga ggagagcatc
     1681 tccgacgcag agaaggtggc catggatctc aaggacccca accgcccccg gttcaccctg
     1741 caggagctgc gggacgtgct gcacgagagg aacgagctca agtccaaggt gttcttgctg
     1801 caggaggagc tggcttacta taagagtgaa gaaatggaag aggaaaaccg aataccccaa
     1861 cccccaccca tcgcccaccc gaggacgtcc ccccagccgg agtcgggcat caagcgactg
     1921 tttagcttct tctcccgaga taagaagcgc ctggccaaca cacagagaaa cgtgcacatc
     1981 caggagtcct ttggacagtg ggcaaacacc caccgcgatg acggttacac agagcaagga
     2041 caggaagccc tgcagcatct gtgaaagggc gaattcgacc cagctttctt gtacaaagtg
     2101 gtgatgagag aagggcgaat tcgaaagtgg tgatcggtac cttaattaat taaggtacca
     2161 ggtaagtgta cccaattcgc cctatagtga gtcgtattac aattcactcg atcgcccttc
     2221 ccaacagttg cgcagcctga atggcgaatg gagatccaat ttttaagtgt ataatgtgtt
     2281 aaactactga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc tcatttcagg
     2341 cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt
     2401 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2461 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2521 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2581 atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2641 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2701 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2761 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2821 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2881 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2941 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3001 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     3061 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3121 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3181 gaagaatcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3301 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3361 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3421 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3481 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3541 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gaacatcagg ggctcgcgcc
     4081 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     4141 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4201 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4261 tattgctgaa gaacttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4321 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4381 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4441 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4501 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4561 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4621 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4681 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4741 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4801 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4861 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4921 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4981 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5041 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5101 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5161 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5221 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5281 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5341 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5401 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5461 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5521 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5581 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5641 gcgttatccc ctgattctgt ggataaccgt attaccgcc
//
