LOCUS       sequence-273271-        6819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing T7 RNA Polymerase in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  TITLE     Lentiviral capsid delivery of SpCas9 RNP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6819)
  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            "Unpublished"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: pCMV-T7 RNA POL
FEATURES             Location/Qualifiers
     source          1..6819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        185..388
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     339..359
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     385..409
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          522..997
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      1065..1074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3805..4199
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(4538..4555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4709..5297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4789..4808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5468..6259,6260..6328))
                     /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     6091..6110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6329..6433)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6503..6522)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        join(6624..6819,1..184)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
       61 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      121 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      181 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      241 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      301 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      361 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      421 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tcccctcgaa
      481 gcttacatgt ggtaccgagc tcggatcctg agaacttcag ggtgagtcta tgggaccctt
      541 gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag gggagaagta
      601 acagggtaca catattgacc aaatcagggt aattttgcat ttgtaatttt aaaaaatgct
      661 ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc
      721 tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag
      781 tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa
      841 attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc
      901 tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt
      961 gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg
     1021 tgtgctggcc catcactttg gcaaagcacg tgagatctga attcgccacc atggacacga
     1081 ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg ttcaacactc
     1141 tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag catgagtctt
     1201 acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa gctggtgagg
     1261 ttgcggataa cgctgccgcc aagcctctca tcactaccct actccctaag atgattgcac
     1321 gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg acagccttcc
     1381 agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag accactctgg
     1441 cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca atcggtcggg
     1501 ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag cacttcaaga
     1561 aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa gcatttatgc
     1621 aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt
     1681 ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc attgagtcaa
     1741 ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac tctgagacta
     1801 tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg ctggctggca
     1861 tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc attactggtg
     1921 gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac agtaagaaag
     1981 cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt aacattgcgc
     2041 aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta atcaccaagt
     2101 ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc ccgatgaaac
     2161 cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct gccgctgctg
     2221 tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc atgcttgagc
     2281 aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg gactggcgcg
     2341 gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc aaaggactgc
     2401 ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg aaaatccacg
     2461 gtgcaaactg tgcgggtgtc gataaggttc cgttccctga gcgcatcaag ttcattgagg
     2521 aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact tggtgggctg
     2581 agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg gtacagcacc
     2641 acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc tctggcatcc
     2701 agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac ttgcttccta
     2761 gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag attctacaag
     2821 cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag aacactggtg
     2881 aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg ctggcttacg
     2941 gtgttactcg cagtgtgact aagcgttcag tcatgacgct ggcttacggg tccaaagagt
     3001 tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat tccggcaagg
     3061 gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg atttgggaat
     3121 ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag tctgctgcta
     3181 agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc aagcgttgcg
     3241 ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag aagcctattc
     3301 agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc attaacacca
     3361 acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct aactttgtac
     3421 acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag aagtacggaa
     3481 tcgaatcttt tgcactgatt cacgactcct tcggtaccat tccggctgac gctgcgaacc
     3541 tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat gtactggctg
     3601 atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa atgccagcac
     3661 ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc gcgttcgcgt
     3721 aacgccaaat caatacgact ccggatccct gcagctcgag tggtggctgg tgtggctaat
     3781 gccctggccc acaagtatca ctaagctcgc tttcttgctg tccaatttct attaaaggtt
     3841 cctttgttcc ctaagtccaa ctactaaact gggggatatt atgaagggcc ttgagcatct
     3901 ggattctgcc taataaaaaa catttatttt cattgcaatg atgtatttaa attatttctg
     3961 aatattttac taaaaaggga atgtgggagg tcagtgcatt taaaacataa agaaatgaag
     4021 agctagttca aaccttggga aaatacacta tatcttaaac tccatgaaag aaggtgaggc
     4081 tgcaaacagc taatgcacat tggcaacagc ccctgatgcc tatgccttat tcatccctca
     4141 gaaaaggatt caagtagagg cttgatttgg aggttaaagt tttgctatgc tgtattttac
     4201 attacttatt gttttagctg tcctcatgaa tgtcttttca ctacccattt gcttatcctg
     4261 catctctcag ccttgactcc actcagttct cttgcttaga gataccacct ttcccctgaa
     4321 gtgttccttc catgttttac ggcgagatgg tttctcctcg cctggccact cagccttagt
     4381 tgtctctgtt gtcttataga ggtctacttg aagaaggaaa aacagggggc atggtttgac
     4441 tgtcctgtga gcccttcttc cctgcctccc ccactcacag tgacccggaa tccctcgaca
     4501 tggcagtcta gcactagtgc ggccgcagat ctgcttcctc gctcactgac tcgctgcgct
     4561 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4621 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4681 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4741 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4801 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4861 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4921 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4981 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5041 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5101 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5161 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5221 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5881 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtggatccc ctgagggggc
     6481 ccccatgggc tagaggatcc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     6541 tgagcttggc ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc
     6601 atgtccaaca ttaccgccat gttgacattg attattgact agttattaat agtaatcaat
     6661 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6721 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6781 tcccatagta acgccaatag ggactttcca ttgacgtca
//
