LOCUS       sequence-305929-        8612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression vector to generate a hypoxia fate-mapping 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8612)
  AUTHORS   Godet I, Shin YJ, Ju JA, Ye IC, Wang G, Gilkes DM
  TITLE     Fate-mapping post-hypoxic tumor cells reveals a ROS-resistant 
            phenotype that promotes metastasis.
  JOURNAL   Nat Commun. 2019 Oct 24;10(1):4862. doi: 10.1038/s41467-019-12412-1.
  PUBMED    31649238
REFERENCE   2  (bases 1 to 8612)
  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 
            Commun."; date: "2019-10-24"; volume: "10(1):4862. doi"; pages: " 
            10.1038/s41467-019-12412-1"
COMMENT     Sequence Label: 4xHRE-MinTK-CRE
FEATURES             Location/Qualifiers
     source          1..8612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1797..1914
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2064..2095
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2169..2741
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2749..2768
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2796..2814
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2830..2848
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2830..2847
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2883..3911
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     LTR             5162..5395
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5467..5588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5504..5523)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5558..5577
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(5753..5771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5781..5798)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5781..5797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5790..5812)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5939..6394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6026..6045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6236..6257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6420..6524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6525..6593,6594..7385)
                     /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     complement(6743..6762)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7556..8144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8045..8064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8298..8315
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8432..8453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8468..8473,8474..8491,8492..8498)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8506..8522
                     /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)."
     primer_bind     8511..8533
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8530..8546
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8530..8546
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8565..8585
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8567..8585
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggtttt
     1801 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     1861 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttatcgac
     1921 tcagaacacg ccgtgtacgt gcttcacgtg tacgtggcga tcgtgtacgt gctgtctcac
     1981 acagcactct agactactcg tgtacgtgta cacgaagctt cgtagcaatc ggaccgacga
     2041 cgccgggctc gagatctaga ctctagaggg tatataatgg aagctcgaat tccagcttgg
     2101 cattccggta ctgttggtaa aaactaggct tttgcaaaaa gctccggatc gatcctgaga
     2161 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     2221 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     2281 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     2341 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     2401 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     2461 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     2521 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     2581 gaaatattct tattggtaga aacaactaca tcctggtcat catcctgcct ttctctttat
     2641 ggttacaatg atatacactg cttgagatga ggataaaata ctctgagtcc aaaccgggcc
     2701 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     2761 ttattgtgct gtctcatcat tttggcaaag aattgtaata cgactcacta tagggcgaat
     2821 tcaagcttga tttaggtgac actatagaat acaagctact tgttcttttt gcaggatcca
     2881 ccatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga
     2941 gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc
     3001 atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata
     3061 accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg
     3121 cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc
     3181 gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga
     3241 tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca
     3301 ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta
     3361 atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga
     3421 tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa
     3481 cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac
     3541 tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt
     3601 gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg
     3661 ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg
     3721 gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg
     3781 cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata
     3841 ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg
     3901 aagatggcga tggttctcta gaacattaag ctagcctagt gccatttgtt cagtggttcg
     3961 tagggctttc ccccactgtt tggctttcag ttatatggat gatgtggtat tgggggccaa
     4021 gtctgtacag catcttgagt ccctttttac cgctgttacc aattttcttt tgtctttggg
     4081 tatacattta aaccctaaca aaacaaagag atggggttac tctctaaatt ttatgggtta
     4141 tgtcattgga tgttatgggt ccttgccaca agaacacatc atacaaaaaa tcaaagaatg
     4201 ttttagaaaa cttcctatta acaggcctat tgattggaaa gtatgtcaac gaattgtggg
     4261 tcttttgggt tttgctgccc cttttacaca atgtggttat cctgcgttga tgcctttgta
     4321 tgcatgtatt caatctaagc aggctttcac tttctcgcca acttacaagg cctttctgtg
     4381 taaacaatac ctgaaccttt accccgttgc ccggcaacgg ccaggtctgt gccaagtgtt
     4441 tgctgacgca acccccactg gctggggctt ggtcatgggc catcagcgca tgcgtggaac
     4501 cttttcggct cctctgccga tccatactgc ggaactccta gccgcttgtt ttgctcgcag
     4561 caggtctgga gcaaacatta tcgggactga taactctgtt gtcctatccc gcaaatatac
     4621 atcgtttcca tggctgctag gctgtgctgc caactggatc ctgcgcggga cgtcctttgt
     4681 ttacgtcccg tcggcgctga atcctgcgga cgacccttct cggggtcgct tgggactctc
     4741 tcgtcccctt ctccgtctgc cgttccgacc gaccacgggg cgcacctctc tttacgcgga
     4801 ctccccgtct gtgccttctc atctgccgga ccgtgtgcac ttcgcttcac ctctgcacgt
     4861 cgcatggaga ccaccgtgaa cgcccaccaa atattgccca aggtcttaca taagaggact
     4921 cttggactct cagcaatgtc aacgaccgac cttgaggcat acttcaaaga ctgtttgttt
     4981 aaagactggg aggagttggg ggaggagatt aggttaaagg tctttgtact aggaggctgt
     5041 aggcataaat tggtctgcgc accagcacca tggcgcaatc actagagcgg ggtaccttta
     5101 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5161 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     5221 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5281 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5341 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     5401 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     5461 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5521 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5581 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatt ctccgcccca
     5641 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5701 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     5761 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5821 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5881 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     5941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6361 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6421 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6481 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6541 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6601 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6661 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6721 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6781 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6841 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6901 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6961 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7021 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7081 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7141 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7201 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7261 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7321 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7381 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8221 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8281 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8341 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8401 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8461 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8521 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     8581 aaagggaaca aaagctggag ctgcaagctt aa
//
