LOCUS       sequence-305919-        8592 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 8592)
  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 8592)
  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: 3xHRE-MinTK-CRE
FEATURES             Location/Qualifiers
     source          1..8592
                     /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        2013..2044
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2118..2690
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2698..2717
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2745..2763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2779..2797
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2779..2796
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2832..3860
                     /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             5111..5344
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5416..5537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5453..5472)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5507..5526
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5577..5712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5733..5751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5761..5778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5761..5777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5770..5792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5919..6374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6006..6025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6216..6237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6400..6504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6505..6573,6574..7365)
                     /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(6723..6742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7536..8124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8025..8044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8278..8295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8412..8433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8448..8453,8454..8471,8472..8478)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8486..8502
                     /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     8491..8513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8510..8526
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8510..8526
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8545..8565
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8547..8565
                     /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 tacgtggcgc acgaagcttc gtagcaatcg
     1981 gaccgacgac gccgggctcg agatctagac tctagagggt atataatgga agctcgaatt
     2041 ccagcttggc attccggtac tgttggtaaa aactaggctt ttgcaaaaag ctccggatcg
     2101 atcctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     2161 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     2221 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     2281 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     2341 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     2401 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     2461 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     2521 gctggcgtgg aaatattctt attggtagaa acaactacat cctggtcatc atcctgcctt
     2581 tctctttatg gttacaatga tatacactgc ttgagatgag gataaaatac tctgagtcca
     2641 aaccgggccc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     2701 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgtaatac gactcactat
     2761 agggcgaatt caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg
     2821 caggatccac catgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg
     2881 atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt
     2941 tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca
     3001 agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat
     3061 atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca
     3121 tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta
     3181 tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt
     3241 tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg
     3301 atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa
     3361 ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata
     3421 ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg
     3481 taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact
     3541 acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat
     3601 caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg
     3661 atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc
     3721 gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca
     3781 atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc
     3841 gcctgctgga agatggcgat ggttctctag aacattaagc tagcctagtg ccatttgttc
     3901 agtggttcgt agggctttcc cccactgttt ggctttcagt tatatggatg atgtggtatt
     3961 gggggccaag tctgtacagc atcttgagtc cctttttacc gctgttacca attttctttt
     4021 gtctttgggt atacatttaa accctaacaa aacaaagaga tggggttact ctctaaattt
     4081 tatgggttat gtcattggat gttatgggtc cttgccacaa gaacacatca tacaaaaaat
     4141 caaagaatgt tttagaaaac ttcctattaa caggcctatt gattggaaag tatgtcaacg
     4201 aattgtgggt cttttgggtt ttgctgcccc ttttacacaa tgtggttatc ctgcgttgat
     4261 gcctttgtat gcatgtattc aatctaagca ggctttcact ttctcgccaa cttacaaggc
     4321 ctttctgtgt aaacaatacc tgaaccttta ccccgttgcc cggcaacggc caggtctgtg
     4381 ccaagtgttt gctgacgcaa cccccactgg ctggggcttg gtcatgggcc atcagcgcat
     4441 gcgtggaacc ttttcggctc ctctgccgat ccatactgcg gaactcctag ccgcttgttt
     4501 tgctcgcagc aggtctggag caaacattat cgggactgat aactctgttg tcctatcccg
     4561 caaatataca tcgtttccat ggctgctagg ctgtgctgcc aactggatcc tgcgcgggac
     4621 gtcctttgtt tacgtcccgt cggcgctgaa tcctgcggac gacccttctc ggggtcgctt
     4681 gggactctct cgtccccttc tccgtctgcc gttccgaccg accacggggc gcacctctct
     4741 ttacgcggac tccccgtctg tgccttctca tctgccggac cgtgtgcact tcgcttcacc
     4801 tctgcacgtc gcatggagac caccgtgaac gcccaccaaa tattgcccaa ggtcttacat
     4861 aagaggactc ttggactctc agcaatgtca acgaccgacc ttgaggcata cttcaaagac
     4921 tgtttgttta aagactggga ggagttgggg gaggagatta ggttaaaggt ctttgtacta
     4981 ggaggctgta ggcataaatt ggtctgcgca ccagcaccat ggcgcaatca ctagagcggg
     5041 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5101 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     5161 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5221 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5281 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5341 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5401 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5461 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5521 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     5581 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5641 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5701 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     5761 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5821 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5881 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     5941 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6001 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6061 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6121 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6181 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6241 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6301 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6361 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6421 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6481 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6541 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6601 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6661 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6721 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6781 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6841 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6901 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6961 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7021 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7081 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7141 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7201 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7261 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7321 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7381 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7441 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7501 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7561 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7621 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7681 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7741 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7801 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7861 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7921 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7981 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8041 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8101 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8161 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8221 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8281 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8341 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8401 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8461 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8521 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     8581 ctgcaagctt aa
//
