LOCUS       sequence-194514-        8608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector containing C-terminally Halo-tag tagged mouse 
            ATF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8608)
  AUTHORS   Seiichi Oyadomari
  TITLE     Oyadomari Lab Luciferase reporters
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8608)
  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: 
            "unpublished"
COMMENT     Sequence Label: pCDF1-mAtf4-HT-puro
FEATURES             Location/Qualifiers
     source          1..8608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        133..336
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        1479..1682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1679..1703
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2838..2858
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2874..3758
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     promoter        3826..4037
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4050..4318
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             4345..4944
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4345..4364)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4841..4861
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4951..5539
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5004..5024)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5422..5433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5876..5997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5913..5932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5967..5986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6037..6172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6205..6221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6205..6221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6218..6240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6229..6245
                     /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)."
     promoter        complement(join(6253..6259,6260..6277,6278..6283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6298..6319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6436..6453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6607..7195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6687..6706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7366..8157,8158..8226))
                     /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     7989..8008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8227..8331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8399..8417
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8455..8477)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8577..8596
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
       61 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      121 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      181 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      241 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      301 aggcgtgtac ggtgggaggt ctatataagc agagcttgtg aaacttcgag gagtctcttt
      361 gttgaggact tttgagttct cccttgaggc tcccacagat acaataaata tttgagattg
      421 aaccctgtcg agtatctgtg taatcttttt tacctgtgag gtctcggaat ccgggccgag
      481 aacttcgcag ttggcgcccg aacagggact tgattgagag tgattgagga agtgaagcta
      541 gagcaataga aagctgttaa gcagaactcc tgctgaccta aatagggaag cagtagcaga
      601 cgctgctaac agtgagtatc tctagtgaag cagactcgag ctcataatca agtcattgtt
      661 taaaggccca gataaattac atctggtgac tcttcgcgga ccttcaagcc aggagattcg
      721 ccgagggaca gtcaacaagg taggagagat tctacagcaa catggggaat ggacaggggc
      781 gagattggaa aatggccatt aagagatgta gtaatgttgc tgtaggagta ggggggaaga
      841 gtaaaaaatt tggagaaggg aatttcagat gggccattag aatggctaat gtatctacag
      901 gacgagaacc tggtgatata ccagagactt tagatcaact aaggttggtt atttgcgatt
      961 tacaagaaag aagagaaaaa tttggatcta gcaaagaaat tgatatggca attcctgcat
     1021 tgaggagaaa tggtaggcaa tgtggcatgt ctgaaaaaga ggaggaatga tgaagtatct
     1081 cagacttatt ttataaggga gatactgtgc tgagttcttc cctttgagga aggtatgtca
     1141 tatcctagac atagtctcaa ttttaaaaga agaggtagga taggagggat ggccccttat
     1201 gaattattag cacaacaaga atccttaaga atacaagatt atttttctgc aataccacaa
     1261 aaattgcaag cacagtggat ttattataaa gatcaaaaag ataagaaatg gaaaggacca
     1321 atgagagtag aatactgggg acagggatca gtattattaa aggatgaaga gaagggatat
     1381 tttcttataa tcgatactag tattatgccc agtacatgac cttatgggac tttcctactt
     1441 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1501 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1561 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1621 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1681 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1741 gaagattcta gagcgccatg accgagatga gcttcctgaa cagcgaagtg ttggcggggg
     1801 acttgatgtc ccccttcgac cagtcgggtt tgggggctga agaaagccta ggtctcttag
     1861 atgactatct ggaggtggcc aagcacttga aacctcatgg gttctccagc gacaaggcgg
     1921 gctcctcgga atggccggct atggatgatg gcttggccag tgcctcagac accggcaagg
     1981 aggatgcctt ttccgggaca gattggatgt tggagaaaat ggatctgaaa gagtttgact
     2041 tcgatgctct gtttcgaatg gatgacctgg aaaccatgcc agatgagctc ttgaccacgt
     2101 tggatgacac atgtgatctt tttgcccctc tagtccaaga gactaataag gagccccctc
     2161 agacagtgaa cccaattggc catctcccag aaagtttaat aaaagtcgac caggttgccc
     2221 cctttacatt cttgcagcct ttcccctgtt ccccaggggt tctgtcttcc actccagagc
     2281 attcctttag tttagagcta ggcagtgaag ttgatatctc tgaaggagac aggaagcctg
     2341 actctgctgc ttacattact ctaatccctc catgtgtaaa ggaggaagac actccctctg
     2401 acaatgacag tggcatctgt atgagcccag agtcctacct gggctctccc cagcatagcc
     2461 cctccacctc cagggcccca ccagacaatc tgccttctcc aggtggttcc cgtgggtctc
     2521 ctcggcccaa accttatgac ccacctggag ttagtttgac agctaaagtg aagactgaga
     2581 aattggataa gaagctgaaa aagatggagc aaaacaagac agcagccact aggtaccgcc
     2641 agaagaagcg ggctgagcag gaggccctca ctggcgagtg taaggagcta gaaaaaaaga
     2701 atgaggctct gaaagagaag gcagattctc tggccaagga gatccagtat ctgaaagacc
     2761 tgatagaaga ggtccgtaag gcaaggggga agaagagagt tccgtctaga gcccgggcgc
     2821 gccggatcca gatctctgag gatctgtact ttcagagcga taacgatgga tccgaaatcg
     2881 gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg
     2941 tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct
     3001 cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag
     3061 acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg
     3121 tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc
     3181 acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag
     3241 gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg
     3301 cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc
     3361 agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg
     3421 agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct
     3481 tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat
     3541 acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg
     3601 ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg
     3661 tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg
     3721 agatcgcgcg ctggctgtct actctggaga tttccggtag atcttaatta atttaaatga
     3781 attcgcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     3841 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     3901 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     3961 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     4021 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     4081 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     4141 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     4201 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     4261 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     4321 agctgtgacc ggcgcctacg ctagatgacc gagtacaagc ccacggtgcg cctcgccacc
     4381 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4441 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4501 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4561 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4621 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4681 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4741 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4801 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4861 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4921 atgacccgca agcccggtgc ctgagtcgac aatcaacctc tggattacaa aatttgtgaa
     4981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5341 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5461 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5521 ctttgggccg cctccccgcc tggtaccgat gacagagtta gaagatcgct tcaggaagct
     5581 atttggcacg acttctacaa cgggagacag cacagtagat tctgaagatg aacctcctaa
     5641 aaaagaaaaa agggtggact gggatgagta ttggaaccct gaaatcgata gcttccagtg
     5701 ctttgtgaaa cttcgaggag tctctttgtt gaggactttt gagttctccc ttgaggctcc
     5761 cacagataca ataaatattt gagattgaac cctgtcgagt atctgtgtaa tcttttttac
     5821 ctgtgaggtc tcggaatccg ggccgagaac ttcgcagcga gctcattgta ccgcgaactt
     5881 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5941 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6001 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6061 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6121 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6181 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     6241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8341 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     8401 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     8461 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     8521 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     8581 atcagagcag attgtactga gagtgcac
//
