LOCUS       sequence-257598-        8147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  extGluc luciferase for invivo Imaging, NGFR for flourescent 
            detection or separation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8147)
  AUTHORS   Ebinger S, Ozdemir EZ, Ziegenhain C, Tiedt S, Castro Alves C, 
            Grunert M, Dworzak M, Lutz C, Turati VA, Enver T, Horny HP, Sotlar 
            K, Parekh S, Spiekermann K, Hiddemann W, Schepers A, Polzer B, 
            Kirsch S, Hoffmann M, Knapp B, Hasenauer J, Pfeifer H, 
            Panzer-Grumayer R, Enard W, Gires O, Jeremias I
  TITLE     Characterization of Rare, Dormant, and Therapy-Resistant Cells in 
            Acute Lymphoblastic Leukemia.
  JOURNAL   Cancer Cell. 2016 Dec 12;30(6):849-862. doi: 
            10.1016/j.ccell.2016.11.002. Epub 2016 Dec 1.
  PUBMED    27916615
REFERENCE   2  (bases 1 to 8147)
  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"; doi: 
            "10.1016/j.ccell.2016.11.002"; journalName: "Cancer Cell"; date: 
            "2016-12-12- 12"; volume: "30"; issue: "6"; pages: "849-862"
COMMENT     Sequence Label: pCDH-EF1a-extGluc-NGFR
FEATURES             Location/Qualifiers
     source          1..8147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    584..709
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             756..936
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(937..1163)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(1178..1195)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1178..1194)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1187..1209)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1403..1422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1522..1544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1582..1600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1668..1772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1773..1841,1842..2633)
                     /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(1991..2010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2804..3392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3546..3563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3680..3701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3716..3721,3722..3739,3740..3746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3754..3770
                     /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     3759..3781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3778..3794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3778..3794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      3827..3962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4002..4123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4013..4032)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4067..4086
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             4195..4428
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4502..5090
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4608..4619
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5017..5037
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5946..5999)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      6809..6818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             6858..7126
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(7139..7350)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    7393..7509
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7776..7820)
                     /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    join(8005..8147,1..91)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc
       61 tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac
      121 cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata
      181 tttatataat tcacttctcc aattgtccct catatctcct cctccaggtc tgaagatcag
      241 tggccgcttg ctgtgcggtg gtcttacttt tgttttgctc ttcctctatc ttgtctaaag
      301 cttccttggt gtcttttatc tctatccttt gatgcacaca atagagggtt gctactgtat
      361 tatataatga tctaagttct tctgatcctg tctgaaggga tggttgtagc tgtcccagta
      421 tttgtctaca gccttctgat gtttctaaca ggccaggatt aactgcgaat cgttctagct
      481 ccctgcttgc ccatactata tgttttaatt tatatttttt ctttccccct ggccttaacc
      541 gaattttttc ccatcgcgat ctaattctcc cccgcttaat actgacgctc tcgcacccat
      601 ctctctcctt ctagcctccg ctagtcaaaa tttttggcgt actcaccagt cgccgcccct
      661 cgcctcttgc cgtgcgcgct tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt
      721 tccctttcgc tttcaagtcc ctgttcgggc gccactgcta gagattttcc acactgacta
      781 aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac acactacttg
      841 aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag
      901 ctcccaggct cagatctggt ctaaccagag agacccgttt atgtatcgag ctaggcactt
      961 aaatacaata tctctgcaat gcggcaattc agtggttcgt ccaatccatg tcagacccgt
     1021 ctgttgcctt cctaataagg cacgatcgta ccaccttact tccaccaatc ggcatgcacg
     1081 gtgctttttc tctccttgta aggcatgttg ctaactcatc gttaccatgt tgcaagacta
     1141 caagagtatt gcataagact acacgcgtca gcttggcact ggccgtcgtt ttacaacgtc
     1201 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1261 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1321 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1381 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1441 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1501 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1561 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1621 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1741 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1801 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1861 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1921 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1981 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2041 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2101 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2161 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2221 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2281 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2341 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2401 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2461 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2521 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2581 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2641 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2941 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3541 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3601 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3661 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     3721 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     3781 gaaacagcta tgacatgatt acgaatttgg tctctgcaaa agtctaggcc tccaaaaaag
     3841 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
     3901 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     3961 atgggcggag ttaggggcgg gatagctaga gccagacatg ataagataca ttgatgagtt
     4021 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     4081 tattgcttta tttgtaacca ttataagctg caataaacaa gttcctctca ctctctgata
     4141 ttcatttctt tgcaagttat aaatactgaa taataagatg acatgaacta ctactgctag
     4201 agattttcca cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca
     4261 gacgggcaca cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt
     4321 tccctagtta gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac
     4381 aagcaaaaag cagatcttat cttcgttggg agtgaattag cccttccagt cccccctttt
     4441 cttttaaaaa gtggctaaga tctacagctg ccttgtaagt cattggtctt aaaggtacca
     4501 ggcggggagg cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga
     4561 agaggccgca gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa
     4621 gggacgtagc agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa
     4681 ggaaaggacg atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc
     4741 cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg
     4801 aaagcgaaag tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt
     4861 gcgtcagcaa acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca
     4921 taaagagaca gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca
     4981 atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag
     5041 ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttag
     5101 attcagcttc agttccacct cttgaaggct atgtaggcca caaggcccac aaccacagca
     5161 gccaggatgg agcaatagac agggatgagg ttgtcggtgg tgcctcgggt caccacgggc
     5221 tgggagctgc ccatcactgt ggtcaccaca cctgccaccg tgctggctat gaggtcttgt
     5281 tctggaggtg cctcaggctc ctgggtgctg ggggctgtgc tgtccgagcc ctctgggggt
     5341 gtggaccgtg taatccaacg gccagggatc tcctcgcact cggcgtcggc ccagcgtgtg
     5401 cactcgcgga gctggcgctc ggtgtcctcg cacacggtgc agggcaggca cgggtccacg
     5461 tggttggcct cgtcggaata cgtgccgtcg gggcactcct cgcacacggt gttctgcttg
     5521 tcctggcagg agaacacgag gcccgagccc gcctcgcaca cgcggcacgc ctcgcagcgc
     5581 ccagtcgtct catcctggta gtagccgtag gcgcagcggc acacggcgtc gtcggcctcc
     5641 acgcacggcg ccgacatgct ctggagcccc acgcactcgg tgcacggctt gcacggctcg
     5701 gtcgcgctca ccacgtcgga gaacgtcacg ctgtccaggc agggctcaca cacggtctgg
     5761 ttggctccac aaggctgggc cacaccctcg cccaggttgc aggctttgca gcactcaccg
     5821 ctgtgtgtgt acaggcctgt ggggcatgcc tccttggcac ctccaaggga cacccccaga
     5881 agcagcaaca gcagcaggcg cggcccgtcc atggcgcggc cggtggcacc tgcccccata
     5941 tgcatagggc cgggattctc ctccacgtca ccgcatgtta gaagacttcc tctgccctca
     6001 gcggccgcgg atccttgaat ccgcctgtgg ttgcagtaaa gggtgataac cagtgacagg
     6061 agaaggaccc cacaagtccc ggccaggggc gcccagatgt agatatcaca ggcgaagtcc
     6121 agccccctcg tgtgcactgc gccccccgcc gctggccggc acgcctctgg gcgcagggat
     6181 aggggctgcg acgcgatcgt gggcgccggg gttggtggtc gcggcgctgg cgtcgtggtg
     6241 ggtgcggccg ctaagtcacc accggccccc ttgatcttgt ccacctggcc ctggatcttg
     6301 ctggcaaagg tcgcacagcg ttgcggcagc cacttcttga gcaggtcaga acactgcacg
     6361 ttggcaagcc ctttgaggca gccagttgtg cagtccacac acagatcgac ctgtgcgatg
     6421 aactgctcca ttggctccaa gtccttgaac ccaggaatct caggaatgtc cacgatcgcc
     6481 tcgcctatgc cgccctgtgc ggactctttg tcgccttcgt aggtgtggca gcgtcctggg
     6541 atgaacttct tcatcttggg cgtgcacttg atgtgggaca ggcagatcag acagcccctg
     6601 gtgcagccag ctttccgggc attggcttcc atctctttga gcacctccag cggcagcttc
     6661 ttgccgggca acttcccgcg gtcagcatcg agatccgtgg tcgcgaagtt gctggccacg
     6721 gccacgatgt tgaagtcttc gttgttctcg gtgggcttgg catgcaggag aagcgctagg
     6781 ggaagcagta gggcagtcac tgggagagcc atggtggcga attcgctagc tctagagtag
     6841 gcgccggtca cagcttggat ctgtaacggc gcagaacaga aaacgaaaca aagacgtaga
     6901 gttgagcaag cagggtcagg caaagcgtgg agagccggct gagtctaggt aggctccaag
     6961 ggagcgccgg acaaaggccc ggtctcgacc tgagctttaa acttacctag acggcggacg
     7021 cagttcagga ggcaccacag gcgggaggcg gcagaacgcg actcaaccgg cgtggatggc
     7081 ggcctcaggt agggcggcgg gcgcgtgaag gagagatgcg agcccctcga agcttcagct
     7141 gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat
     7201 acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt
     7261 ttaccccgcg ccaccttctc taggcacccg ttcaattgcc gacccctccc cccaacttct
     7321 cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcga tcgcagatcc
     7381 actagtatcg ataaaatttt gaattttgta atttgttttt gtaattcttt agtttgtatg
     7441 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     7501 cttttaaaag ttaaccgata ccgtcgagat ccgttcacta atcgaatgga tctgtctctg
     7561 tctctctctc caccttcttc ttctattcct tcgggcctgt cgggtcccct cggggttggg
     7621 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     7681 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     7741 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     7801 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     7861 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     7921 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     7981 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     8041 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     8101 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccaga
//
