LOCUS       sequence-169541-        6573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Late1-O3-33-myc-PH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  AUTHORS   Votteler J, Ogohara C, Yi S, Hsia Y, Nattermann U, Belnap DM, King 
            NP, Sundquist WI
  TITLE     Designed proteins induce the formation of nanocage-containing 
            extracellular vesicles.
  JOURNAL   Nature. 2016 Dec 8;540(7632):292-295. doi: 10.1038/nature20607. Epub
            2016 Nov 30.
  PUBMED    27919066
REFERENCE   2  (bases 1 to 6573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature20607"; journalName: "Nature"; date: "2016-12-8- 8"; 
            volume: "540"; issue: "7632"; pages: "292-295"
COMMENT     Sequence Label: pCMV-EPN-51
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        31..410
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        411..614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     565..585
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     659..678
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        659..677
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1886..1905)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(1960..1977)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1966..2190
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2236..2664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2323..2342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2673..2693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2678..3007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2858..2993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2920..2939
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3074..3868
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3128..3147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3738..3757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4042..4163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4079..4098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4133..4152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4212..4228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4212..4228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4225..4247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4236..4252
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4260..4266,4267..4284,4285..4290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4305..4326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4443..4460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4614..5202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4694..4713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5373..6164,6165..6233))
                     /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"
     primer_bind     5996..6015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6234..6338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6413..6432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
       61 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      181 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      241 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      301 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      361 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      421 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      481 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      541 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      601 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
      661 atacgactca ctatagggag acccaagctg gctagcgttt aaacgggccc tctagaatta
      721 tcgatccgga ggtaccatgc tgcagtctcg tccggaaccg accgcaccgc cggaagaatc
      781 tttccgttct ggtgttgaaa ccaccacgcc accgcaaaaa caggagccga tcgacaaaga
      841 actgtacccg ctgacctctc tgcgttctct gttcggtaac gacccgtctt ctcagagcca
      901 ggcaattggt attctggaac tgaccagcat tgcagcaggt atggaactgg gtgatgcaat
      961 gctgaaaagc gcgaacgtgg atctgctggt gagcaaaacc attagcccgg gtaaatttct
     1021 gctgatgctg ggcggtgata ttggtgcgat tcagcaggcg attgaaaccg gtaccagcca
     1081 ggcgggcgaa ctgctggtgg atagcctggt gctggcaaac attcatccga gcgtgctgcc
     1141 ggcgattagc ggtctgaaca gcgtggataa gcgtcaggcg gtgggtattg tggaaacctg
     1201 gagcgtggcg gcatgtatta gcgcagcgga tcgtgcggtg aagggtagca acgtgaccct
     1261 ggtgcgtgtg cacatggcat ttggtattgg cggtaaatgt tatatggtgg tggcgggcga
     1321 tgtgagcgat gtggcactgg cggtgaccgt ggcaagcagc agcgcaggcg catacggtct
     1381 gctggtgtat gcaagcctga ttccgcgtcc gcatgaagca atgtggcgtc agatggtgga
     1441 aggccagaag ctgatctccg aagaagacct gcacggtctg caggacgacc cggacctgca
     1501 agcactgctc aagggcagcc agctgctgaa agttaaatct tcttcttggc gtcgtgaacg
     1561 tttctacaaa ctccaggagg actgcaaaac catctggcag gaatcccgta aagttatgcg
     1621 ttctccggaa tctcagctgt tctctatcga agacatccag gaagttcgta tgggtcaccg
     1681 taccgaaggt ctggaaaaat tcgcgcgtga catcccggaa gaccgttgct tctccatcgt
     1741 tttcaaagac cagcgtaaca ccctggacct gatcgcgccg tctccggcgg acgcgcagca
     1801 ctgggttcag ggtctgcgta aaatcatcca ccactctggt tctatggacc aacgtcagaa
     1861 atgactcgag aagcttgcta gcagatcttt ttccctctgc caaaaattat ggggacatca
     1921 tgaagcccct tgagcactta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt
     1981 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2041 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2101 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2161 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2221 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2641 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     2701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2761 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2821 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2881 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2941 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3001 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3061 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3121 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3181 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3241 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3301 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3361 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3421 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3481 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3541 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3601 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3661 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     3721 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     3781 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     3841 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     3901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4021 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4081 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4141 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4381 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4441 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4501 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4561 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4621 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4681 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4741 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4801 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4861 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4921 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4981 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5041 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5101 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5161 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5221 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5281 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5341 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5401 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5461 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5521 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5581 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5641 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5701 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5761 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5821 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5881 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5941 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6001 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6061 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6121 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6181 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6241 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6301 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6361 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6421 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6481 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     6541 agaatctgct tagggttagg cgttttgcgc tgc
//
