LOCUS       sequence-261580-        8707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CXCR4-com, NLS-com-GNCR1-P2a-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8707)
  AUTHORS   Foight GW, Wang Z, Wei CT, Jr Greisen P, Warner KM, 
            Cunningham-Bryant D, Park K, Brunette TJ, Sheffler W, Baker D, Maly 
            DJ
  TITLE     Multi-input chemical control of protein dimerization for programming
            graded cellular responses.
  JOURNAL   Nat Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8.
  PUBMED    31501561
REFERENCE   2  (bases 1 to 8707)
  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: "Nat 
            Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8."
COMMENT     Sequence Label: GWF314
FEATURES             Location/Qualifiers
     source          1..8707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1447..1680
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1865..1909
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2207..2324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2577..2610)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2614..2634
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     terminator      2825..2844
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     enhancer        2976..3279
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3280..3483
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3520..3540
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4453..4482
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4492..4548
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4549..5247
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     protein_bind    complement(5270..5303)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5359..5947
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5412..5432)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5830..5841)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5950..5966)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5951..5967)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6476..6656
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6718..7306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6798..6817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7477..8268,8269..8337))
                     /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     8100..8119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8338..8442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8517..8536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
     1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2341 acagcagaga tccagtttgg ttagtaccgg gcccgctcta gagatccgac gcgccatctc
     2401 taggcccgcg ccggccccct cgcacggact tgtgggagaa gctcggctac tcccctgccc
     2461 cggttaattt gcatataata tttcctagta actatagagg cttaatgtgc gataaaagac
     2521 agataatctg ttctttttaa tactagctac attttacatg ataggcttgg atttctataa
     2581 cttcgtatag catacattat acgaagttat aaacagcaca aaaggaaact caccctaact
     2641 gtaaagtaat tgtgtgtttt gagactataa gtatcccttg gagaaccacc ttgttggcag
     2701 acgcgaggaa ggagggcgcg tttaagagct atgctggaaa cagcatagca agtttaaata
     2761 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcctgaa tgcctgcgag
     2821 catctttttt tgttttttat gtctctgcag agttcggtct cgagtactag gatccattag
     2881 gcggccgcgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     2941 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     3001 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     3061 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     3121 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     3181 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     3241 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     3301 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3361 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     3421 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3481 gctggtttag tgaaccgtca gatccgctag cccaccatgc ccaaaaagaa aagaaaagtg
     3541 ggtagtatga aatcaattcg ctgtaaaaac tgcaacaaac tgttatttaa ggcggatagt
     3601 tttgatcaca ttgaaatcag gtgtccgcgt tgcaaacgtc acatcataat gctgaatgcc
     3661 tgcgagcatc ccacggagaa acattgtggg aaaagagaaa aaatcacgca ttctgacgaa
     3721 accgtgcgtt atggctccgg tagcggctct ggtgatatcg agaagctttg caaaaaagct
     3781 gaagaagagg caaaggaggc gcaagagaaa gctgatgagt tgcggcaacg gcaccccgac
     3841 agccaggctg ctgaggatgc tgaggacttg gcaaatctgg cggtggccgc tgtcttgacg
     3901 gcttgcttgt tggcccaaga acatccaaac gctgacattg caaagctttg cattaaggcc
     3961 gcgtccgaag cggctgaagc agcttctaag gcagccgagc tggcgcaacg gcatcctgac
     4021 tcccaagcgg cgagggacgc tataaaactc gcatcacaag cagccagagc tgtaatcctc
     4081 gctattatgc tcgcagccga aaacccaaac gctgatattg ccaaactgtg tataaaggcg
     4141 gctagtgaag ccgcagaagc tgcaagcaaa gcagcagaac ttgcccaacg ccaccccgac
     4201 tcccaggcgg ctagagatgc tataaaactt gcttctcagg ctgcggaggc ggtagaacga
     4261 gcgatctggc tggcagcaga gaaccccaac gctgacattg ccaaaaagtg tatcaaggcg
     4321 gcttccgaag ccgccgagga ggccagcaaa gcggcggaag aggcacagag acatccggac
     4381 tcacagaaag cacgcgacga gattaaagag gctagtcaaa aagcagagga ggtaaaagaa
     4441 cgctgtaagt ccgagcaaaa actcatatct gaggaggact tgggaagcgg agctactaac
     4501 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctag cgagctgatt
     4561 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag
     4621 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
     4681 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc
     4741 agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct
     4801 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     4861 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     4921 ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag
     4981 acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg
     5041 ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag
     5101 aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc
     5161 aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct
     5221 agcaaactgg ggcacaagct taattgataa gaattcgtcg agggacctaa taacttcgta
     5281 tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta ggtacaattc
     5341 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5401 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5461 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5521 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5581 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5641 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5701 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5761 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5821 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5881 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5941 tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag caatcacaag
     6001 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     6061 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     6121 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     6181 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     6241 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     6301 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     6361 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     6421 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     6481 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6541 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     6601 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat
     6661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6721 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     7141 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     7201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     7321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     7381 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     7441 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     7501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     7561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     7621 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7921 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     8101 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     8161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     8221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     8281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     8341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     8401 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     8461 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     8521 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     8581 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     8641 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     8701 acgcgtt
//
