LOCUS       2xmyc-RAB10(T73A        5070 bp DNA     circular SYN 04-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5070)
  AUTHORS   BonetPonce lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Viewer 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      670..679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             679..708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDL"
     CDS             724..753
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /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"
                     /note="color: #31849b; direction: RIGHT"
     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"
                     /note="color: #31849b; direction: RIGHT"
     primer_bind     853..875
                     /label=Insert.FOR
                     /note="color: black; sequence: atggcgaagaagacgtacgacct"
     primer_bind     complement(1053..1085)
                     /label=Phosphonull- T73A
                     /note="color: black; sequence: 
                     taggaggttgtgatggcgtgaaatcgctcctgg"
     primer_bind     complement(1435..1455)
                     /label=Insert.REV
                     /note="color: black; sequence: TCAGCAGCATTTGCTCTTCCA"
     protein_bind    complement(1469..1493)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
                     /note="color: #31849b; direction: LEFT"
     promoter        complement(1572..1590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    1864..1985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      complement(1992..2447)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2474..2578
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     promoter        2580..2937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2788..2923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             2972..3766
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3998..4045
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     rep_origin      4374..4962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
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 ttatggcgaa gaagacgtac gacctgcttt tcaagctgct cctgatcggg
      901 gattccggag tggggaagac ctgcgtcctt tttcgttttt cggatgatgc cttcaatact
      961 acctttattt ccaccatagg aatagacttc aagatcaaaa cagttgaatt acaaggaaag
     1021 aagatcaagc tacagatatg ggatacagca ggccaggagc gatttcacgc catcacaacc
     1081 tcctactaca gaggcgcaat gggtatcatg ctagtatatg acatcaccaa tggtaaaagt
     1141 tttgaaaaca tcagcaaatg gcttagaaac atagatgagc atgccaatga agatgtggaa
     1201 agaatgttac taggaaacaa gtgtgatatg gacgacaaaa gagttgtacc taaaggaaaa
     1261 ggagaacaga ttgcaaggga gcatggtatt aggttttttg agactagtgc aaaagcaaat
     1321 ataaacatcg aaaaggcgtt cctcacgtta gctgaagata tccttcgaaa gacccctgta
     1381 aaagagccca acagtgaaaa tgtagatatc agcagtggag gaggcgtgac aggctggaag
     1441 agcaaatgct gctgaaaggg cgaattcgac ccagctttct tgtacaaagt ggtgatgaga
     1501 gaagggcgaa ttcgaaagtg gtgatcggta ccttaattaa ttaaggtacc aggtaagtgt
     1561 acccaattcg ccctatagtg agtcgtatta caattcactc gatcgccctt cccaacagtt
     1621 gcgcagcctg aatggcgaat ggagatccaa tttttaagtg tataatgtgt taaactactg
     1681 attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag gcccctcagt
     1741 cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt tttacttgct
     1801 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1861 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1921 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1981 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2041 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2101 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2161 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2221 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2281 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2341 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2401 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2461 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2521 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatcc
     2581 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2641 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2701 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2761 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2821 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     2881 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     2941 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3001 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3061 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3121 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3181 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3241 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3301 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3361 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3421 cggtcttgtc gatcaggatg atctggacga agaacatcag gggctcgcgc cagccgaact
     3481 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3541 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3601 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3661 agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3721 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3781 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3841 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3901 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     3961 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4021 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4081 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4141 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4201 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4261 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4321 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4381 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4441 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4501 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4561 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4621 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4681 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4741 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4801 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4861 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4921 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4981 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5041 cctgattctg tggataaccg tattaccgcc
//
