LOCUS       sequence-305935-        8684 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 8684)
  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 8684)
  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: 9xHRE-MinTK-CRE
FEATURES             Location/Qualifiers
     source          1..8684
                     /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        2105..2136
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2210..2782
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2790..2809
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2837..2855
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2871..2889
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2871..2888
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2924..3952
                     /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             5203..5436
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5508..5629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5545..5564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5599..5618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5669..5804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5825..5843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5853..5870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5853..5869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5862..5884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6011..6466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6098..6117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6308..6329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6492..6596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6597..6665,6666..7457)
                     /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(6815..6834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7628..8216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8117..8136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8370..8387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8504..8525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8540..8545,8546..8563,8564..8570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8578..8594
                     /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     8583..8605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8602..8618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8602..8618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8637..8657
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8639..8657
                     /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 cgacgtgtac gtgggctacg tgtacgtgct
     2041 gtctcacaca gcacgaagct tcgtagcaat cggaccgacg acgccgggct cgagatctag
     2101 actctagagg gtatataatg gaagctcgaa ttccagcttg gcattccggt actgttggta
     2161 aaaactaggc ttttgcaaaa agctccggat cgatcctgag aacttcaggg tgagtttggg
     2221 gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa
     2281 agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg
     2341 ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt
     2401 ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt
     2461 cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa
     2521 tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg
     2581 ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag
     2641 aaacaactac atcctggtca tcatcctgcc tttctcttta tggttacaat gatatacact
     2701 gcttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca
     2761 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     2821 ttttggcaaa gaattgtaat acgactcact atagggcgaa ttcaagcttg atttaggtga
     2881 cactatagaa tacaagctac ttgttctttt tgcaggatcc accatgtcca atttactgac
     2941 cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa
     3001 cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct
     3061 gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc
     3121 agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa
     3181 aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg
     3241 accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga
     3301 tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg
     3361 ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat
     3421 tgcttataac accctgttac gtatagccga aattgccagg atcagggtta aagatatctc
     3481 acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac
     3541 cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc
     3601 cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg
     3661 tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga
     3721 agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg
     3781 gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat
     3841 accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg
     3901 taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg atggttctct
     3961 agaacattaa gctagcctag tgccatttgt tcagtggttc gtagggcttt cccccactgt
     4021 ttggctttca gttatatgga tgatgtggta ttgggggcca agtctgtaca gcatcttgag
     4081 tcccttttta ccgctgttac caattttctt ttgtctttgg gtatacattt aaaccctaac
     4141 aaaacaaaga gatggggtta ctctctaaat tttatgggtt atgtcattgg atgttatggg
     4201 tccttgccac aagaacacat catacaaaaa atcaaagaat gttttagaaa acttcctatt
     4261 aacaggccta ttgattggaa agtatgtcaa cgaattgtgg gtcttttggg ttttgctgcc
     4321 ccttttacac aatgtggtta tcctgcgttg atgcctttgt atgcatgtat tcaatctaag
     4381 caggctttca ctttctcgcc aacttacaag gcctttctgt gtaaacaata cctgaacctt
     4441 taccccgttg cccggcaacg gccaggtctg tgccaagtgt ttgctgacgc aacccccact
     4501 ggctggggct tggtcatggg ccatcagcgc atgcgtggaa ccttttcggc tcctctgccg
     4561 atccatactg cggaactcct agccgcttgt tttgctcgca gcaggtctgg agcaaacatt
     4621 atcgggactg ataactctgt tgtcctatcc cgcaaatata catcgtttcc atggctgcta
     4681 ggctgtgctg ccaactggat cctgcgcggg acgtcctttg tttacgtccc gtcggcgctg
     4741 aatcctgcgg acgacccttc tcggggtcgc ttgggactct ctcgtcccct tctccgtctg
     4801 ccgttccgac cgaccacggg gcgcacctct ctttacgcgg actccccgtc tgtgccttct
     4861 catctgccgg accgtgtgca cttcgcttca cctctgcacg tcgcatggag accaccgtga
     4921 acgcccacca aatattgccc aaggtcttac ataagaggac tcttggactc tcagcaatgt
     4981 caacgaccga ccttgaggca tacttcaaag actgtttgtt taaagactgg gaggagttgg
     5041 gggaggagat taggttaaag gtctttgtac taggaggctg taggcataaa ttggtctgcg
     5101 caccagcacc atggcgcaat cactagagcg gggtaccttt aagaccaatg acttacaagg
     5161 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     5221 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5281 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5341 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5401 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     5461 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     5521 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5581 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     5641 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5701 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5761 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     5821 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     5881 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5941 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6001 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6061 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6121 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6181 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6421 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     6481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8281 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8341 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8401 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8461 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8521 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     8581 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     8641 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaa
//
