LOCUS       shSRP19_9_in_pLK        8816 bp    DNA     circular SYN 07-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8816)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    176..197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(212..217,218..235,236..242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    250..266
                     /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     274..290
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..329
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        357..583
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             584..764
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    811..936
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1429..1662
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1847..1891
                     /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"
     promoter        2180..2274
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    2228..2246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2256..2274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    2281..2334
                     /label=shSRP19_9
     misc_feature    2375..2492
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2541..3051
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     intron          3060..3632
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        3687..3705
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3716..4336
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     misc_feature    4371..4944
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4964..5563
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             5691..5924
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5996..6117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6157..6292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6313..6331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6341..6357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6499..6954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6980..7084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7085..7153,7154..7945)
                     /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"
     rep_origin      8116..8704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
       61 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      121 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      181 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      241 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      301 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta
      361 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
      421 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
      481 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
      541 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
      601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
      841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
      901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
      961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1261 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     1321 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1381 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     1441 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     1501 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     1561 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     1621 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     1681 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1741 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     1801 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     1861 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     1921 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     1981 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     2041 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     2101 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga
     2161 ctagcctcga gcggccgcaa tatttgcatg tcgctatgtg ttctgggaaa tcaccataaa
     2221 cgtgaaatcc ctatcagtga tagagactta taagttccct atcagtgata gagacaccgg
     2281 atgaatggag acttctaatt tctcgagaaa ttagaagtct ccattcattt tttgaattct
     2341 cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2401 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2461 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2521 cactttggcc gcggctcgag ggggttgggg ttgcgccttt tccaaggcag ccctgggttt
     2581 gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg
     2641 tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg
     2701 ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg
     2761 gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc agacggacag
     2821 cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag cggctgctca
     2881 gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg
     2941 gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc
     3001 acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca gggggatctg
     3061 tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc atgttatatg
     3121 gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg tatcaccatg
     3181 gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca ttgtctcctc
     3241 ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa
     3301 tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa tcactttttt
     3361 ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga acaattgtta
     3421 taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt ggaaatattc
     3481 ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta tggttacaat
     3541 gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc ccctctgcta
     3601 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     3661 tgtctcatca ttttggcaaa gaattgtaat acgactcact atagggcgaa ttgatatgtc
     3721 tagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
     3781 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
     3841 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag
     3901 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa
     3961 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg
     4021 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca
     4081 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt
     4141 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac
     4201 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc
     4261 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa
     4321 atgtgaaagt gggtccgcgt acagcggctc ccgggagttc tagggatctg cccctctccc
     4381 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc
     4441 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc
     4501 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt
     4561 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct
     4621 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa
     4681 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt
     4741 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag
     4801 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt
     4861 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     4921 ttcctttgaa aaacacgatg ataaggatcc accggagctt accatgaccg agtacaagcc
     4981 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     5041 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     5101 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     5161 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     5221 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     5281 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     5341 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     5401 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     5461 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     5521 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     5581 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg gtacctttaa gaccaatgac
     5641 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5701 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     5761 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     5821 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     5881 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     5941 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6121 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6181 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6241 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     6301 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     6361 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6421 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6481 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6541 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6601 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6661 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6721 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6781 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6841 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6901 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     6961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7081 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7201 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7261 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7321 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7381 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7501 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7621 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7681 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7801 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7921 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7981 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     8281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8761 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttg
//
