LOCUS       sequence-422659-        6636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PH-Late1-I3-01-myc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  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 6636)
  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-24
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        123..326
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     277..297
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     371..390
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        371..389
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1607..1636
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1661..1680)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     complement(1735..1752)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1741..1965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2011..2439
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2308..2329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2448..2468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2453..2782
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2633..2768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2695..2714
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2849..3643
                     /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(2903..2922)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3513..3532
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3817..3938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3854..3873)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3908..3927
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3987..4003)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3987..4003)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4000..4022)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4011..4027
                     /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(4035..4041,4042..4059,4060..4065))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4080..4101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4218..4235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4389..4977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4469..4488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5148..5939,5940..6008))
                     /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     5771..5790
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6009..6113)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6188..6207)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6379..6636,1..122)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
       61 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      121 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      181 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      241 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      301 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      361 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacgggc
      421 cctctagaat tatcgatccg gaggtaccat gcacggtctg caggacgacc cggacctgca
      481 agcactgctc aagggcagcc agctgctgaa agttaaatct tcttcttggc gtcgtgaacg
      541 tttctacaaa ctccaggagg actgcaaaac catctggcag gaatcccgta aagttatgcg
      601 ttctccggaa tctcagctgt tctctatcga agacatccag gaagttcgta tgggtcaccg
      661 taccgaaggt ctggaaaaat tcgcgcgtga catcccggaa gaccgttgct tctccatcgt
      721 tttcaaagac cagcgtaaca ccctggacct gatcgcgccg tctccggcgg acgcgcagca
      781 ctgggttcag ggtctgcgta aaatcatcca ccactctggt tctatggacc aacgtcagaa
      841 actgcagtct cgtccggaac cgaccgcacc gccggaagaa tctttccgtt ctggtgttga
      901 aaccaccacg ccaccgcaaa aacaggagcc gatcgacaaa gaactgtacc cgctgacctc
      961 tctgcgttct ctgttcggta acgacccgtc ttctcagaag atcgaagagc tgttcaagaa
     1021 acacaagatc gttgccgtgc tgcgtgccaa tagtgtggaa gaagcgaaaa agaaagcgct
     1081 ggcggttttc ctgggcggcg ttcatctgat tgaaattacc tttaccgtgc cggatgcgga
     1141 taccgtgatt aaggaactga gctttctgaa ggaaatgggc gcgattattg gtgcgggcac
     1201 cgtgaccagc gtggagcagt gccgtaaagc ggtggaaagt ggcgccgaat tcattgtgag
     1261 tccgcacctg gacgaggaaa ttagccaatt ttgcaaggag aagggtgtgt tctatatgcc
     1321 aggcgttatg accccgaccg aactggtgaa agccatgaaa ctgggccata ccatcttaaa
     1381 actgtttccg ggtgaggtgg tgggtccgca gtttgttaaa gcgatgaaag gtccgtttcc
     1441 gaatgtgaaa tttgtgccaa ccggcggtgt taatctggac aatgtgtgcg aatggttcaa
     1501 agcgggcgtg ctggccgtgg gcgtgggcag cgcgttagtg aaaggcaccc cggtggaagt
     1561 ggcggaaaag gccaaggcgt tcgttgagaa gattcgtggc tgcaccgaac agaagctgat
     1621 ctccgaagaa gacctgtgac tcgagaagct tgctagcaga tctttttccc tctgccaaaa
     1681 attatgggga catcatgaag ccccttgagc acttaagttt aaaccgctga tcagcctcga
     1741 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1801 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1861 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1921 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     1981 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2401 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2461 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2521 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2581 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2641 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2701 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2761 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     2821 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2881 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2941 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3001 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3061 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3121 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3181 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3241 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3301 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3361 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3421 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3481 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3541 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3601 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3661 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3721 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3781 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3841 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3901 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3961 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4021 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4081 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4141 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4201 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4801 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5041 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5101 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5161 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5221 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5281 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5341 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5401 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5461 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5521 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5581 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5641 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5701 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5761 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5821 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5881 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5941 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6001 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6061 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6121 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     6181 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     6241 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     6301 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     6361 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6421 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6481 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6541 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6601 cccacttggc agtacatcaa gtgtatcata tgccaa
//
