LOCUS       sequence-257354-        7109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral control shRNA vector with puromycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  AUTHORS   Huang S, Sinicrope FA
  TITLE     Sorafenib inhibits STAT3 activation to enhance TRAIL-mediated 
            apoptosis in human pancreatic cancer cells.
  JOURNAL   Mol Cancer Ther. 2010 Mar . 9(3):742-50.
  PUBMED    20197401
REFERENCE   2  (bases 1 to 7109)
  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: "Mol Cancer 
            Ther. 2010 Mar . 9(3):742-50."
COMMENT     Sequence Label: pSIH1-puro-control shRNA
FEATURES             Location/Qualifiers
     source          1..7109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     384..401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    518..539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(554..559,560..577,578..584)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    592..608
                     /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     597..619
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     616..632
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     616..632
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      665..800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(851..870)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     905..924
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             1033..1213
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1310..1524)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(1370..1389)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    1636..2224
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1742..1753
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2151..2171
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2231..2830)
                     /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(2314..2334)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2811..2830
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(2879..2903)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2900..3103)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2929..2949)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    3193..3309
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(3576..3620)
                     /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    3805..4038
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4531..4656
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4703..4883
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(4884..5110)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(5125..5142)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5125..5141)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5134..5156)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5350..5369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5469..5491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5529..5547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5615..5719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5720..5788,5789..6580)
                     /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(5938..5957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6751..7109,1..230)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
       61 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      241 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      301 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      361 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      421 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      481 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      541 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      601 gataacaatt tcacacagga aacagctatg acatgattac gaatttggtc tctgcaaaag
      661 tctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
      721 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
      781 ggcggagtta ggggcgggat gggcggagtt aggggcggga tagctagagc cagacatgat
      841 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
      901 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
      961 tcctctcact ctctgatatt catttctttg caagttataa atactgaata ataagatgac
     1021 atgaactact actgctagag attttccaca ctgactgaaa gggtctgagg gatctctagt
     1081 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     1141 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     1201 accagagaga cccagtagaa gcaaaaagca gaatcgaaga attcaaaaaa aatagcgact
     1261 aaacacatca atctcttgaa ttgatgtgtt tagtcgctat tggatccaag tggtctcata
     1321 cagaacttat aagattccca aatccaaaga catttcacgt ttatggtgat ttcccagaac
     1381 acatagcgac atgcaaatat tgcagggcgc cactcccctg tccctcacag ccatcttcct
     1441 gccagggcgc acgcgcgctg ggtgttcccg cctagtgaca ctgggcccgc gattccttgg
     1501 agcgggttga tgacgtcagc gttccaattc ttgacatcgt tgggagtgaa ttagcccttc
     1561 cagtcccccc ttttctttta aaaagtggct aagatctaca gctgccttgt aagtcattgg
     1621 tcttaaaggt accaggcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa
     1681 ggcggagacg cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg
     1741 gattgagggc cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacac
     1801 aggcgagcag ccaaggaaag gacgatgatt tccccgacaa caccacggaa ttgtcagtgc
     1861 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
     1921 caatagggag ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc
     1981 aaccagtggg ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg
     2041 gccacaactc ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag
     2101 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     2161 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     2221 gattgtcgac tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca
     2281 cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg
     2341 cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca
     2401 cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga
     2461 accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg
     2521 ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg
     2581 cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca
     2641 ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct
     2701 cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg
     2761 cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
     2821 actcggtcat ctagaatctt ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     2881 tctgacggtt cactaaacga gctctgctta tataaacctc ccaccgtaca cgcctaccgc
     2941 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     3001 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     3061 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     3121 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     3181 actagtatcg ataaaatttt gaattttgta atttgttttt gtaattcttt agtttgtatg
     3241 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     3301 cttttaaaag ttaaccgata ccgtcgagat ccgttcacta atcgaatgga tctgtctctg
     3361 tctctctctc caccttcttc ttctattcct tcgggcctgt cgggtcccct cggggttggg
     3421 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     3481 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     3541 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     3601 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     3661 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     3721 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     3781 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     3841 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     3901 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     3961 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     4021 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     4081 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     4141 cttctccaat tgtccctcat atctcctcct ccaggtctga agatcagcgg ccgcttgctg
     4201 tgcggtggtc ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc
     4261 ttttatctct atcctttgat gcacacaata gagggttgct actgtattat ataatgatct
     4321 aagttcttct gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc
     4381 ttctgatgtt tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca
     4441 tactatatgt tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca
     4501 tcgcgatcta attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta
     4561 gcctccgcta gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt
     4621 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
     4681 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
     4741 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     4801 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     4861 atctggtcta accagagaga cccgtttatg tatcgagcta ggcacttaaa tacaatatct
     4921 ctgcaatgcg gcaattcagt ggttcgtcca atccatgtca gacccgtctg ttgccttcct
     4981 aataaggcac gatcgtacca ccttacttcc accaatcggc atgcacggtg ctttttctct
     5041 ccttgtaagg catgttgcta actcatcgtt accatgttgc aagactacaa gagtattgca
     5101 taagactaca cgcgtcagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5161 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5221 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5281 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     5341 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5401 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5461 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5521 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5581 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5641 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5701 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5761 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5821 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5881 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5941 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6001 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6061 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6121 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6181 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6241 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6301 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6361 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6421 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6481 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6541 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6601 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6661 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7081 gtgcacacag cccagcttgg agcgaacga
//
