LOCUS       Exported                5989 bp ds-DNA     circular SYN 04-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5989)
  AUTHORS   BonetPonce lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 4, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5989
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(600..619)
                     /label=Vector.REV
     primer_bind     605..643
                     /label=Fragment.FOR
     regulatory      614..619
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(1295..1336)
                     /label=Fragment.REV
     primer_bind     1322..1352
                     /label=Vector.FOR
     primer_bind     1322..1352
                     /label=Vector.REV
     primer_bind     1322..1336
                     /label=Fragment.FOR (1)
     protein_bind    1346..1370
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1384..1403
                     /label=Fragment.FOR (1)
     primer_bind     1385..1403
                     /label=RILPL1_F
     primer_bind     1385..1403
                     /label=Insert.FOR
     primer_bind     complement(2572..2596)
                     /label=Fragment.REV (1)
     primer_bind     complement(2572..2596)
                     /label=Insert.REV
     primer_bind     complement(2572..2593)
                     /label=RILPL1_R
     protein_bind    complement(2610..2634)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2784..2905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2912..3367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3394..3498
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3500..3857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3708..3843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3892..4686
                     /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    4918..4965
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5294..5882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggtctcaaa gggagaggag gaaaacatgg ctagtacacc
      661 atttaaattt caacttaaag gaaccatcaa tggcaaatcg tttaccgttg aaggcgaagg
      721 tgaagggaac tcacatgaag gttctcataa aggaaaatat gtttgtacaa gtggaaaact
      781 accgatgtca tgggcagcac ttgggacaac ctttggttat ggaatgaaat attataccaa
      841 atatcctagt ggactgaaga actggtttcg tgaagtaatg cccggaggct ttacctacga
      901 tcgtcatatt caatataaag gcgatgggag tatccatgca aaacaccaac actttatgaa
      961 aaatgggact tatcacaaca ttgtagaatt taccggtcag gattttaaag aaaatagtcc
     1021 agtcttaact ggagatatga atgtctcatt accgaatgaa gtcccacaaa tacccagaga
     1081 tgatggagta gaatgcccag tgaccttgct ttatccttta ttatcggata aatcaaaata
     1141 cgtcgaggct caccaatata caatctgcaa gcctcttcat aatcaaccag cacctgatgt
     1201 cccatatcac tggattcgta aacaatacac acaaagcaaa gatgatgccg aggaacgcga
     1261 tcatatctgt caatcagaga ctctcgaagc acacttaaag ggcatggacg agctgtataa
     1321 gtccggactc agatctcgag atcaaacaag tttgtacaaa aaagcaggct ccgaattcgc
     1381 ccttatggag gaggagcggg ggtcggcgct ggcggccgag tcggcgctgg agaagaacgt
     1441 ggccgagctg accgtcatgg acgtgtacga catcgcgtcg cttgtgggcc acgagttcga
     1501 gcgggtcatt gaccagcacg gctgcgaggc catcgcgcgc ctcatgccca aggtcgtgcg
     1561 cgtcctggag atcctggagg tgctggtcag ccgccaccac gtcgcgcccg agctggacga
     1621 gctgcgcctg gagctggacc gcctgcgcct ggagaggatg gaccgcatcg agaaggagcg
     1681 caagcaccag aaggagctgg agctggtgga ggatgtgtgg cgaggggagg cgcaggacct
     1741 cctctcccag atcgcccagc tgcaggagga gaacaagcag ctcatgacca acctctccca
     1801 caaggatgtc aacttctcag aggaggagtt ccagaagcat gaaggcatgt cagagcggga
     1861 gcgacaggtg atgaagaagc tgaaggaggt ggtggacaaa caacgcgacg agatccgcgc
     1921 caaggacagg gagctgggcc tgaaaaatga ggacgttgag gctttacagc agcagcagac
     1981 acggctgatg aagatcaacc atgaccttcg gcaccgggtc acggtggtgg aggcccaggg
     2041 gaaagccctg atcgaacaga aggtggagct ggaggcagac ctgcagacca aggagcagga
     2101 gatgggcagc ctgcgagcag agctggggaa gttgcgagag aggctgcagg gggagcacag
     2161 ccagaatggg gaggaggagc ctgagacgga gccggtggga gaggagagca tctccgacgc
     2221 agagaaggtg gccatggatc tcaaggaccc caaccgcccc cggttcaccc tgcaggagct
     2281 gcgggacgtg ctgcacgaga ggaacgagct caagtccaag gtgttcttgc tgcaggagga
     2341 gctggcttac tataagagtg aagaaatgga agaggaaaac cgaatacccc aacccccacc
     2401 catcgcccac ccgaggacgt ccccccagcc ggagtcgggc atcaagcgac tgtttagctt
     2461 cttctcccga gataagaagc gcctggccaa cacacagaga aacgtgcaca tccaggagtc
     2521 ctttggacag tgggcaaaca cccaccgcga tgacggttac acagagcaag gacaggaagc
     2581 cctgcagcat ctgtgaaagg gcgaattcga cccagctttc ttgtacaaag tggttcgatg
     2641 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     2701 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2761 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2821 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2881 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2941 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3001 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3061 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3121 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3181 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3241 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3301 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3361 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3421 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3481 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3661 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3721 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3781 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3841 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3901 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3961 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4021 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4081 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4141 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4201 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4261 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4321 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4381 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4441 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4501 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4561 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4621 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4681 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4741 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4801 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4861 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4921 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4981 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5041 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5101 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5161 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5221 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5401 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5461 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5761 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5821 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5881 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5941 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgc
//
