LOCUS       sequence-393124-        7978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 3X HIV NES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7978)
  AUTHORS   Price RM, Budzynski MA, Shen J, Mitchell JE, Kwan JZJ, Teves SS
  TITLE     Heat shock transcription factors demonstrate a distinct mode of 
            interaction with mitotic chromosomes.
  JOURNAL   Nucleic Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304.
  PUBMED    37114996
REFERENCE   2  (bases 1 to 7978)
  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"; journalName: "Nucleic 
            Acids Res. 2023 Apr 28:gkad304. doi: 10.1093/nar/gkad304."
COMMENT     Sequence Label: NES-Halo
FEATURES             Location/Qualifiers
     source          1..7978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     481..500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(719..823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(849..1304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(986..1007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1198..1217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1431..1453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1445..1462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1446..1462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1592..1654
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2107..2318
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2331..2599
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      2625..2634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2634..2699
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2709..2735
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2745..2771
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2781..2807
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2814..3704
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYRVPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             3717..3737
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     misc_feature    3768..4340
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3935..3952)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4162..4181
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4349..5143
                     /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
                     VLFVKTDLSGALNELQDEAARLSWLAATGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4403..4422)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5013..5032
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5203..5223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    5265..5386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5302..5321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5356..5375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6150..6184
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6675..6691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6675..6691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6688..6710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6699..6715
                     /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(6723..6729,6730..6747,6748..6753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6768..6789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6906..6923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7077..7665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7157..7176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7836..7978,1..649,650..718))
                     /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"
ORIGIN
        1 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
       61 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      841 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
      901 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      961 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1021 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1081 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1141 agggagcccc cgatttagag cttgacgggg aaagccggcg atcgtggcga ggaaggaagg
     1201 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     1261 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     1321 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     1381 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     1441 acgttgtaaa acgacggcca gtgagcgcgc ctcgttcatt cacgtttttg aacccgtgga
     1501 ggacgggcag actcgcggtg caaatgtgtt ttacagcgtg atggagcaga tgaagatgct
     1561 cgacacgctg cagaacacgc agctagatta accctagaaa gataatcata ttgtgacgta
     1621 cgttaaagat aatcatgtgt aaaattgacg catgtgtttt atcggtctgt atatcgaggt
     1681 ttatttatta atttgaatag atattaagtt ttattatatt tacacttaca tactaataat
     1741 aaattcaaca aacaatttat ttatgtttat ttatttatta aaaaaaacaa aaactcaaaa
     1801 tttcttctat aaagtaacaa aacttttatg agggacagcc cccccccaaa gcccccaggg
     1861 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
     1921 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
     1981 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
     2041 cctgggggga tacggggaaa aggcctccac ggccaaggat ctgcgatcgc tccggtgccc
     2101 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     2161 attgaacggg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     2221 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     2281 acgttctttt tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca
     2341 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
     2401 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
     2461 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
     2521 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
     2581 ttctgcgccg ttacagatcc aagctgtgac cggcgcctac tctagccacc atggactaca
     2641 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagc
     2701 ttcttcaact gcctccactt gaaagactga cactggatct gcaactcccc ccccttgaaa
     2761 gactgacact ggatcttcag ctgcctccac tcgaaagact gacactgggc tccatggcag
     2821 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
     2881 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
     2941 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
     3001 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
     3061 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
     3121 tcattcacga ctggggctcc gctctgggct tccactgggc caagcgcaat ccagagcgcg
     3181 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     3241 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     3301 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     3361 aagtcgagat ggaccattac cgcgtgccgt tcctgaatcc tgttgaccgc gagccactgt
     3421 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     3481 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     3541 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     3601 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     3661 gcagcgagat cgcccgctgg ctgtcgacgc tcgagatttc cggcgagcca accactgagg
     3721 atctgtactt tcagagcgat aactccggac tcagatccgc ggccgcgccc ctctccctcc
     3781 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3841 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3901 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3961 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     4021 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     4081 ccacgtgtat aggatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     4141 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4201 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     4261 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4321 ccttgaaaaa cacgataata cctccggaat gattgaacaa gatggattgc acgcaggttc
     4381 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4441 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4501 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4561 cgcgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4621 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4681 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4741 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4801 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4861 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4921 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4981 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5041 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5101 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagtcgaca atcaacctct
     5161 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5221 atgtggatac gctgctttaa tgcctttgta tcatgcgtta actaaacttg tttattgcag
     5281 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5341 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggaact
     5401 gactcaaatg atgtcaatta gtctatcaga agctcatctg gtctcccttc cgggggacaa
     5461 gacatccctg tttaatattt aaacagcagt gttcccaaac tgggttctta tatcccttgc
     5521 tctggtcaac caggttgcag ggtttcctgt cctcacagga acgaagtccc taaagaaaca
     5581 gtggcagcca ggtttagccc cggaattgac tggattcctt ttttagggcc cattggtatg
     5641 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     5701 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggcac
     5761 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     5821 agcgggggag ggacgtaatt acatccctgg gggctttggg gggggggctg tccccgatat
     5881 ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat
     5941 ataattatcg tatgagttaa atcttaaaag tcacgtaaaa ggtaatcatg cgtcattttg
     6001 actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc
     6061 acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
     6121 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
     6181 agggttaatc tagctgcatc aggatcatat cgtcgggtct tttttccggc tcagtcatcg
     6241 cccaagctgg cgctatctgg gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt
     6301 tgaagcggca tggaaagagt ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa
     6361 acgcacgttt accatgatga ttcgggaagg tgtggccacg cacgccttta acggtgaact
     6421 gttcgttcag gccacctggg ataccagttc gtcgcggctt ttccggacac agttccggat
     6481 ggtcagcccg aagcgcatca gcaacccgaa caataccggc gacagccgga actgccgtgc
     6541 cggtgtgcag attaatgaca gcggtgcggc gctgggatat tacgtcagcg aggacgggta
     6601 tcctggctgg atgccgcaga aatggacatg gataccccgt gagttacccg gcgggcgcgc
     6661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     6721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     6781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc ggggagcgtg
     7261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     7681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     7741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     7801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgag
//
