LOCUS       sequence-267190-        6817 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 6817)
  TITLE     Lentiviral capsid delivery of SpCas9 RNP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6817)
  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            "Unpublished"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..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        541..744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     695..715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..765
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          878..1353
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      1421..1430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4160..4553
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     primer_bind     complement(4892..4909)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5063..5651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5143..5162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5822..6613,6614..6682))
                     /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     6445..6464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6683..6787)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatcccctg agggggcccc catgggctag aggatccggc ctcggcctct gcataaataa
       61 aaaaaattag tcagccatga gcttggccca ttgcatacgt tgtatccata tcataatatg
      121 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      601 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      661 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      721 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      781 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccc ctcgaagctt
      841 acatgtggta ccgagctcgg atcctgagaa cttcagggtg agtctatggg acccttgatg
      901 ttttctttcc ccttcttttc tatggttaag ttcatgtcat aggaagggga gaagtaacag
      961 ggtacacata ttgaccaaat cagggtaatt ttgcatttgt aattttaaaa aatgctttct
     1021 tcttttaata tacttttttg tttatcttat ttctaatact ttccctaatc tctttctttc
     1081 agggcaataa tgatacaatg tatcatgcct ctttgcacca ttctaaagaa taacagtgat
     1141 aatttctggg ttaaggcaat agcaatattt ctgcatataa atatttctgc atataaattg
     1201 taactgatgt aagaggtttc atattgctaa tagcagctac aatccagcta ccattctgct
     1261 tttattttat ggttgggata aggctggatt attctgagtc caagctaggc ccttttgcta
     1321 atcatgttca tacctcttat cttcctccca cagctcctgg gcaacgtgct ggtctgtgtg
     1381 ctggcccatc actttggcaa agcacgtgag atctgaattc gccaccatgg acacgattaa
     1441 catcgctaag aacgacttct ctgacatcga actggctgct atcccgttca acactctggc
     1501 tgaccattac ggtgagcgtt tagctcgcga acagttggcc cttgagcatc agtcttacga
     1561 gatgggtgaa gcacgcttcc gcaagatgtt tgagcgtcaa cttaaagctg gtgaggttgc
     1621 ggataacgct gccgccaagc ctctcatcac taccctactc cctaagatga ttgcacgcat
     1681 caacgactgg tttgagcaag tgaaagctaa gcgcggcaag cgcccgacag ccttccagtt
     1741 cctgcaagaa atcaagccgg aagccgtagc gtacatcacc attaagacca ctctggcttg
     1801 cctaaccagt gctcacaata caaccgttca ggctgtagca agcgcaatcg gtcgggccat
     1861 tgaggacgag gctcgcttcg gtcgtatccg tgaccttgaa gcattgcact tcaagaaaaa
     1921 cgttgaggaa caactcaaca agcgcgtagg gcacgtctac aagaaagcat ttatgcaagt
     1981 tgtcgaggct gacatgctct ctaagggtct actcggtggc gaggcgtggt cttcgtggca
     2041 taaggaagac tctattcatg taggagatcg ctgcatcgag atgctcattg actcaaccgg
     2101 aatggttagc ttacaccgcc aaaatgctgg cgtagtaggt caagactctg agactatcga
     2161 actcgcacct gaatacgctg aggctatcgc aacccgtgca ggtgcgctgg ctggcatctc
     2221 tccgatgttc caaccttgcg tagttcctcc taagccgtgg actggcatta ctggtggtgg
     2281 ctattgggct aacgctcgtc gtcctctggc gctggtgcgt actcacagta agaaagcact
     2341 gatgcgctac gaagacgttt acatgcctga ggtgtacaaa gcgattaaca ttgcgcaaaa
     2401 caccgcatgg aaaatcaaca agaaagtcct agcggtcgcc aacgtaatca ccaagtggaa
     2461 gcattgtccg gtcgaggaca tccctgcgat tgagcgtgaa gaactcccga tgaaaccgga
     2521 agacatcgac atgaatcctg aggctctcac cgcgtggaaa cgtgctgccg ctgctgtgta
     2581 ccgcaagaca aggctcgcaa gtctcgccgt atcagccttg agttcatgct tgagcaagcc
     2641 aataagtttg ctaaccataa ggccatctgg ttcccttaca acatggactg gcgccgttcg
     2701 tgtttacgct gtgtcaatgt tcaacccgca aggtaacgat atgaccaaag gacgtcttac
     2761 gctggcgaaa ggtaaaccaa tcggtaagga aggttactac tggctgaaaa tccacggtgc
     2821 aaactgtgcg ggtgtcgata aggtttcgtt tcctgagcgc atcaagttca ttgaggaaaa
     2881 ccacgagaac atcatggctt gcgctaagtc tccactggag aacacttggt gggctgagca
     2941 agattctccg ttctgcttcc ttgcgttctg ctttgagtac gctggggtac agcaccacgg
     3001 cctgagctat aactgctccc ttccgctggc gtttgacggg tcttgctctg gcatccagca
     3061 cttctccgcg atgctccgag atgaggatgg tcgtcgcgcg gttaacttgc ttcctagtca
     3121 aaccgttcag gacatcatcg ggattgttgc taagaaagtc aacgagattc tgcaagcaga
     3181 cgcaatcaat gggaccgata acgaagtact taccgtgacc gatgagaaca ctggtgaaat
     3241 ctctgagaaa gtcaagctgg gcactaaggc actggctggt caatggctgg cttacggtgt
     3301 tactcgcagt gtgactaagc gttcagtcat gacgctggct tacgggtcca aagagttcgg
     3361 cttccgtcaa caagtgctgg aagataccat tcagccagct attgattccg gcaagggtct
     3421 gatgttcact cagccgaatc aggctgctgg atacatggct aagctgattt gggaatccgt
     3481 gagcgtgacg gtggtagctg cggttgaagc aatgaactgg cttaagtctg ctgctaagct
     3541 gctggctgct gaggtcaaag ataagaagac tggagagatt cttcgcaagc gttgcgctgt
     3601 gcattgggta actcctgatg gtttccctgt gtggcaggaa tacaagaagc ctattcagac
     3661 gcgcttgaac ctgatgttcc tcggtcagtt ccgcttacag cctaccatta acaccaacaa
     3721 agatagcgag attgatgcac acaaacagga gtctggtatc gctcctaact ttgtacacag
     3781 ccaagacggt agccaccttc gtaagactgt agtgtgggca cacgagaagt acggaatcga
     3841 atcttttgca ctgattcacg actccttcgg taccattccg gctgacgctg cgaacctgtt
     3901 caaagcagtg cgcgaaacta tggttgacac atatgagtct tgtgatgtac tggctgattt
     3961 ctacgaccag ttcgctgacc agttgcacga gtctcaattg gacaaaatgc cagcacttcc
     4021 ggctaaaggt aacttgaacc tccgtgacat cttagagtcg gacttcgcgt tcgcgtaacg
     4081 ccaaatcaat acgactcgga tccctgcagc tcgagtggtg gctggtgtgg ctaatgccct
     4141 ggcccacaag tttcactaag ctcgcttcct tgctgtccaa tttctattaa aggttccttg
     4201 gttccctaag tccaactact aaactggggg atattatgaa gggccttgag catctggatt
     4261 ctgcctaata aaaaacattt attttcattg caatgatgta tttaaattat ttctgaatat
     4321 tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa tgaagagcta
     4381 gttcaaacct tgggaaaata cactatatct taaactccat gaaagaaggt gaggctgcaa
     4441 acagctaatg cacattggca acagccctga tgcctatgcc ttattcatcc ctcagaaaag
     4501 gattcaagta gaggcttgat ttggaggtta aagtttggct atgctgtatt ttacattact
     4561 tattgtttta gctgtcctca tgaatgtctt ttcactaccc atttgcttat cctgcatctc
     4621 tcagccttga ctccactcag ttctcttgct tagagatacc acctttcccc tgaagtgttc
     4681 cttccatgtt ttacggcgag atggtttctc ctcgcctggc cactcagcct tagttgtctc
     4741 tgttgtctta tagaggtcta cttgaagaag gaaaaacagg gggcatggtt tgactgtcct
     4801 gtgagccctt cttccctgcc tcccccactc acagtgaccc ggaatccctc gacatggcag
     4861 tctagcacta gtgcggccgc agatctgctt cctcgctcac tgactcgctg cgctcggtcg
     4921 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4981 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5221 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     5281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5461 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5521 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5821 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5881 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5941 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6001 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6061 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6781 ttccgcgcac atttccccga aaagtgccac ctgacgt
//
