LOCUS       sequence-305934-        8631 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 8631)
  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 8631)
  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: 5xHRE-MinTK-CRE
FEATURES             Location/Qualifiers
     source          1..8631
                     /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        2052..2083
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2157..2729
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2737..2756
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2784..2802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2818..2836
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2818..2835
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2871..3899
                     /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             5150..5383
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5455..5576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5492..5511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5546..5565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5616..5751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5772..5790)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5800..5817)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5800..5816)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5809..5831)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5958..6413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6045..6064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6255..6276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6439..6543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6544..6612,6613..7404)
                     /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(6762..6781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7575..8163
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8064..8083
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8317..8334
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8451..8472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8487..8492,8493..8510,8511..8517)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8525..8541
                     /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     8530..8552
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8549..8565
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8549..8565
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8584..8604
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8586..8604
                     /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 agactactca cgaagcttcg tagcaatcgg accgacgacg ccgggctcga
     2041 gatctagact ctagagggta tataatggaa gctcgaattc cagcttggca ttccggtact
     2101 gttggtaaaa actaggcttt tgcaaaaagc tccggatcga tcctgagaac ttcagggtga
     2161 gtttggggac ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag
     2221 ggggcaaagt tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac
     2281 cctcatgata attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta
     2341 ttttcttttc attttctgta actttttcgt taaactttag cttgcatttg taacgaattt
     2401 ttaaattcac ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc
     2461 aaggcaatca gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa
     2521 ttaaatgata aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta
     2581 ttggtagaaa caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat
     2641 atacactgct tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc
     2701 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     2761 ctcatcattt tggcaaagaa ttgtaatacg actcactata gggcgaattc aagcttgatt
     2821 taggtgacac tatagaatac aagctacttg ttctttttgc aggatccacc atgtccaatt
     2881 tactgaccgt acaccaaaat ttgcctgcat taccggtcga tgcaacgagt gatgaggttc
     2941 gcaagaacct gatggacatg ttcagggatc gccaggcgtt ttctgagcat acctggaaaa
     3001 tgcttctgtc cgtttgccgg tcgtgggcgg catggtgcaa gttgaataac cggaaatggt
     3061 ttcccgcaga acctgaagat gttcgcgatt atcttctata tcttcaggcg cgcggtctgg
     3121 cagtaaaaac tatccagcaa catttgggcc agctaaacat gcttcatcgt cggtccgggc
     3181 tgccacgacc aagtgacagc aatgctgttt cactggttat gcggcggatc cgaaaagaaa
     3241 acgttgatgc cggtgaacgt gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc
     3301 aggttcgttc actcatggaa aatagcgatc gctgccagga tatacgtaat ctggcatttc
     3361 tggggattgc ttataacacc ctgttacgta tagccgaaat tgccaggatc agggttaaag
     3421 atatctcacg tactgacggt gggagaatgt taatccatat tggcagaacg aaaacgctgg
     3481 ttagcaccgc aggtgtagag aaggcactta gcctgggggt aactaaactg gtcgagcgat
     3541 ggatttccgt ctctggtgta gctgatgatc cgaataacta cctgttttgc cgggtcagaa
     3601 aaaatggtgt tgccgcgcca tctgccacca gccagctatc aactcgcgcc ctggaaggga
     3661 tttttgaagc aactcatcga ttgatttacg gcgctaagga tgactctggt cagagatacc
     3721 tggcctggtc tggacacagt gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag
     3781 tttcaatacc ggagatcatg caagctggtg gctggaccaa tgtaaatatt gtcatgaact
     3841 atatccgtaa cctggatagt gaaacagggg caatggtgcg cctgctggaa gatggcgatg
     3901 gttctctaga acattaagct agcctagtgc catttgttca gtggttcgta gggctttccc
     3961 ccactgtttg gctttcagtt atatggatga tgtggtattg ggggccaagt ctgtacagca
     4021 tcttgagtcc ctttttaccg ctgttaccaa ttttcttttg tctttgggta tacatttaaa
     4081 ccctaacaaa acaaagagat ggggttactc tctaaatttt atgggttatg tcattggatg
     4141 ttatgggtcc ttgccacaag aacacatcat acaaaaaatc aaagaatgtt ttagaaaact
     4201 tcctattaac aggcctattg attggaaagt atgtcaacga attgtgggtc ttttgggttt
     4261 tgctgcccct tttacacaat gtggttatcc tgcgttgatg cctttgtatg catgtattca
     4321 atctaagcag gctttcactt tctcgccaac ttacaaggcc tttctgtgta aacaatacct
     4381 gaacctttac cccgttgccc ggcaacggcc aggtctgtgc caagtgtttg ctgacgcaac
     4441 ccccactggc tggggcttgg tcatgggcca tcagcgcatg cgtggaacct tttcggctcc
     4501 tctgccgatc catactgcgg aactcctagc cgcttgtttt gctcgcagca ggtctggagc
     4561 aaacattatc gggactgata actctgttgt cctatcccgc aaatatacat cgtttccatg
     4621 gctgctaggc tgtgctgcca actggatcct gcgcgggacg tcctttgttt acgtcccgtc
     4681 ggcgctgaat cctgcggacg acccttctcg gggtcgcttg ggactctctc gtccccttct
     4741 ccgtctgccg ttccgaccga ccacggggcg cacctctctt tacgcggact ccccgtctgt
     4801 gccttctcat ctgccggacc gtgtgcactt cgcttcacct ctgcacgtcg catggagacc
     4861 accgtgaacg cccaccaaat attgcccaag gtcttacata agaggactct tggactctca
     4921 gcaatgtcaa cgaccgacct tgaggcatac ttcaaagact gtttgtttaa agactgggag
     4981 gagttggggg aggagattag gttaaaggtc tttgtactag gaggctgtag gcataaattg
     5041 gtctgcgcac cagcaccatg gcgcaatcac tagagcgggg tacctttaag accaatgact
     5101 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5161 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     5221 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5281 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     5341 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     5401 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     5461 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     5521 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5581 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5641 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5701 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     5761 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     5821 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5881 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5941 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6001 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6061 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6121 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6181 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6241 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     6301 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     6361 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     6421 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6481 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6541 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6601 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6661 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6721 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6781 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6841 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6901 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6961 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7021 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7081 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7141 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7201 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7261 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7321 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7381 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7441 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7501 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7561 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7621 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     7681 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     7741 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     7801 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     7861 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7921 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7981 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8041 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8101 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8161 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8221 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8281 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8341 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8401 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8461 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8521 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8581 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta a
//
