LOCUS       pLenti_SFNpromot        8249 bp DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        167..205
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             310..987
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     misc_feature    989..1577
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(1460..1471)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        1591..2090
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2111..2710
                     /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             2848..3081
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3153..3274
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3314..3449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3470..3488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3498..3514)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3656..4111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4137..4241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4242..4310,4311..5102)
                     /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      5273..5861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6149..6170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6185..6190,6191..6208,6209..6215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6223..6239
                     /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     6247..6263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6284..6302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6330..6556
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6557..6737
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6784..6909
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7402..7635
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7820..7864
                     /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    8131..8247
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggatccacgg gataccccag gggctctcca gggaatctcc ggggatactc cagggggttt
       61 ccggggaatc ccccgggagt ttcctgggaa tttcccggga tttccccggg gcatcccggg
      121 gactctcctg ggattttcca gggacattcc tgggactttc ctgcgcggta ggcgtgtacg
      181 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      241 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctct cgacattcgt
      301 gccaccatga tggcgagtag cgaagacgtt atcaaagagt tcatgcgttt caaagttcgt
      361 atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga aggtcgtccg
      421 tacgaaggta cccagaccgc taaactgaaa gttaccaaag gtggtccgct gccgttcgct
      481 tgggacatcc tgtccccgca gttccagtac ggttccaaag cttacgttaa acacccggct
      541 gacatcccgg actacctgaa actgtccttc ccggaaggtt tcaaatggga acgtgttatg
      601 aacttcgaag acggtggtgt tgttaccgtt acccaggact cctccctgca agacggtgag
      661 ttcatctaca aagttaaact gcgtggtacc aacttcccgt ccgacggtcc ggttatgcag
      721 aaaaaaacca tgggttggga agcttccacc gaacgtatgt acccggaaga cggtgctctg
      781 aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg gtcactacga cgctgaagtt
      841 aaaaccacct acatggctaa aaaaccggtt cagctgccgg gtgcttacaa aaccgacatc
      901 aaactggaca tcacctccca caacgaagac tacaccatcg ttgaacagta cgaacgtgct
      961 gaaggtcgtc actccaccgg tgcttaacaa tcaacctctg gattacaaaa tttgtgaaag
     1021 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     1081 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     1141 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     1201 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     1261 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     1321 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     1381 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     1441 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     1501 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     1561 ttgggccgcc tccccgcctg gaattctacc gggtagggga ggcgcttttc ccaaggcagt
     1621 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     1681 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     1741 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     1801 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     1861 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     1921 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     1981 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     2041 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     2101 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2161 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2221 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     2281 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     2341 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2401 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     2461 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     2521 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     2581 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     2641 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     2701 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     2761 tgcatctcga gggcccggta cctttaagac caatgactta caaggcagct gtagatctta
     2821 gccacttttt aaaagaaaag gggggactgg aagggctagc tcactcccaa cgaagacaag
     2881 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     2941 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     3001 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     3061 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     3121 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     3181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     3301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3421 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     3481 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3541 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     3601 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     3661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3721 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3841 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3961 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4021 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4081 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4921 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5221 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5401 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5641 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6121 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6181 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6241 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     6301 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
     6361 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
     6421 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
     6481 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
     6541 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6601 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6661 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6721 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     6781 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     6841 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     6901 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     6961 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     7021 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     7081 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     7141 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     7201 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     7261 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     7321 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     7381 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     7441 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     7501 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     7561 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     7621 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     7681 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     7741 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     7801 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     7861 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     7921 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     7981 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     8041 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     8101 agtgaacgga tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta
     8161 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     8221 aaaacaaatt acaaaattca aaattttat
//
