LOCUS       sequence-305922-        8576 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 8576)
  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 8576)
  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: 1xHRE-MinTK-CRE
FEATURES             Location/Qualifiers
     source          1..8576
                     /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        1997..2028
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     intron          2102..2674
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2682..2701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2729..2747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2763..2781
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2763..2780
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2816..3844
                     /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             5095..5328
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5400..5521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5437..5456)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5491..5510
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5561..5696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(5717..5735)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5745..5762)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5745..5761)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5754..5776)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5903..6358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5990..6009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6200..6221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6384..6488
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6489..6557,6558..7349)
                     /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(6707..6726)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7520..8108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8009..8028
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8262..8279
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8396..8417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8432..8437,8438..8455,8456..8462)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8470..8486
                     /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     8475..8497
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8494..8510
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8494..8510
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8529..8549
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8531..8549
                     /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 gctcacgaag cttcgtagca atcggaccga cgacgccggg
     1981 ctcgagatct agactctaga gggtatataa tggaagctcg aattccagct tggcattccg
     2041 gtactgttgg taaaaactag gcttttgcaa aaagctccgg atcgatcctg agaacttcag
     2101 ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat tcatgttata
     2161 tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct tgtatcacca
     2221 tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac cattgtctcc
     2281 tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc atttgtaacg
     2341 aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct aatcactttt
     2401 ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga gaacaattgt
     2461 tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc gtggaaatat
     2521 tcttattggt agaaacaact acatcctggt catcatcctg cctttctctt tatggttaca
     2581 atgatataca ctgcttgaga tgaggataaa atactctgag tccaaaccgg gcccctctgc
     2641 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     2701 gctgtctcat cattttggca aagaattgta atacgactca ctatagggcg aattcaagct
     2761 tgatttaggt gacactatag aatacaagct acttgttctt tttgcaggat ccaccatgtc
     2821 caatttactg accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga
     2881 ggttcgcaag aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg
     2941 gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa
     3001 atggtttccc gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg
     3061 tctggcagta aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc
     3121 cgggctgcca cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa
     3181 agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt
     3241 cgaccaggtt cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc
     3301 atttctgggg attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt
     3361 taaagatatc tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac
     3421 gctggttagc accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga
     3481 gcgatggatt tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt
     3541 cagaaaaaat ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga
     3601 agggattttt gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag
     3661 atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc
     3721 tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat
     3781 gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg
     3841 cgatggttct ctagaacatt aagctagcct agtgccattt gttcagtggt tcgtagggct
     3901 ttcccccact gtttggcttt cagttatatg gatgatgtgg tattgggggc caagtctgta
     3961 cagcatcttg agtccctttt taccgctgtt accaattttc ttttgtcttt gggtatacat
     4021 ttaaacccta acaaaacaaa gagatggggt tactctctaa attttatggg ttatgtcatt
     4081 ggatgttatg ggtccttgcc acaagaacac atcatacaaa aaatcaaaga atgttttaga
     4141 aaacttccta ttaacaggcc tattgattgg aaagtatgtc aacgaattgt gggtcttttg
     4201 ggttttgctg ccccttttac acaatgtggt tatcctgcgt tgatgccttt gtatgcatgt
     4261 attcaatcta agcaggcttt cactttctcg ccaacttaca aggcctttct gtgtaaacaa
     4321 tacctgaacc tttaccccgt tgcccggcaa cggccaggtc tgtgccaagt gtttgctgac
     4381 gcaaccccca ctggctgggg cttggtcatg ggccatcagc gcatgcgtgg aaccttttcg
     4441 gctcctctgc cgatccatac tgcggaactc ctagccgctt gttttgctcg cagcaggtct
     4501 ggagcaaaca ttatcgggac tgataactct gttgtcctat cccgcaaata tacatcgttt
     4561 ccatggctgc taggctgtgc tgccaactgg atcctgcgcg ggacgtcctt tgtttacgtc
     4621 ccgtcggcgc tgaatcctgc ggacgaccct tctcggggtc gcttgggact ctctcgtccc
     4681 cttctccgtc tgccgttccg accgaccacg gggcgcacct ctctttacgc ggactccccg
     4741 tctgtgcctt ctcatctgcc ggaccgtgtg cacttcgctt cacctctgca cgtcgcatgg
     4801 agaccaccgt gaacgcccac caaatattgc ccaaggtctt acataagagg actcttggac
     4861 tctcagcaat gtcaacgacc gaccttgagg catacttcaa agactgtttg tttaaagact
     4921 gggaggagtt gggggaggag attaggttaa aggtctttgt actaggaggc tgtaggcata
     4981 aattggtctg cgcaccagca ccatggcgca atcactagag cggggtacct ttaagaccaa
     5041 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     5101 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     5161 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5221 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5281 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     5341 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     5401 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5461 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5521 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     5581 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5641 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5701 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     5761 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     5821 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     5881 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6301 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     6361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6721 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7021 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8161 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8221 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8281 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     8341 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     8401 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     8461 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     8521 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaa
//
