LOCUS       Exported                7413 bp ds-DNA     circular SYN 23-SEP-2022
DEFINITION  3rd generation lentiviral plasmid expressing mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    pLV-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  TITLE     Tsoulfas Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7413
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    6..27
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(144..161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(315..903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(395..414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1074..1934)
                     /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     1697..1716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1935..2039)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2065..2520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2202..2223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2414..2433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2647..2669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2661..2678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2662..2678
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2688..2707
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2688..2706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2727..2862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2781..2800)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2902..3023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2913..2932)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2967..2986
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3095..3328
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3415..4003
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3521..3532
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3930..3950
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4010..4720)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4288..4307)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4557..4575
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      4717..4726
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(4791..4815)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4812..5015)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4841..4861)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5016..5319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    5342..5458
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5725..5769)
                     /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    5954..6187
                     /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    6680..6805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6852..7032
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7033..7259)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(7287..7307)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7287..7305)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7326..7342)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7326..7342)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7339..7361)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7350..7366
                     /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)."
     promoter        complement(7374..7404)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
       61 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
      121 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
      181 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
      241 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      301 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
      361 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
      421 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
      481 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
      541 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
      601 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
      661 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
      721 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      781 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      841 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      901 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
      961 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1021 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1081 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1141 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1201 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1261 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1321 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     1381 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     1441 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     1501 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     1561 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     1621 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1681 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1741 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     1801 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1861 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1921 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     1981 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2041 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     2101 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2161 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2221 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2281 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     2341 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     2401 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     2461 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2521 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2581 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2641 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     2701 tatagggcga attgggtacg tccctaggcc tccaaaaaag cctcctcact acttctggaa
     2761 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     2821 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     2881 gatagctaga gccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2941 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     3001 ttataagctg caataaacaa gttcctctca ctctctgata ttcatttctt tgcaagttat
     3061 aaatactgaa taataagatg acatgaacta ctactgctag agattttcca cactgactaa
     3121 aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca cactacttga
     3181 agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta gccagagagc
     3241 tcccaggctc agatctggtc taaccagaga gacccagtac aagcaaaaag cagatcttgt
     3301 cttcgttggg agtgaattag cccttccagt cccccctttt cttttaaaaa gtggctaaga
     3361 tctacagctg ccttgtaagt cattggtctt aaaggtaccg agctcgaatt ccaggcgggg
     3421 aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
     3481 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
     3541 agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc catggaaagg
     3601 acgtcagctt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
     3661 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
     3721 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
     3781 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
     3841 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     3901 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     3961 taccagtcaa tctttcacaa attttgtaat ccagaggttg attgtcgact tacttgtaca
     4021 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     4081 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     4141 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     4201 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     4261 gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     4321 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     4381 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     4441 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     4501 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     4561 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     4621 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     4681 tgatgatggc catgttatcc tcctcgccct tgctcaccat ggtggcggcg gatcctctag
     4741 agtcggtgtc ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg
     4801 ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     4861 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     4921 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     4981 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     5041 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     5101 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     5161 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     5221 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     5281 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccca agcttatcga
     5341 taaaattttg aattttgtaa tttgtttttg taattcttta gtttgtatgt ctgttgctat
     5401 tatgtctact attctttccc ctgcactgta ccccccaatc cccccttttc ttttaaaagt
     5461 taaccgatac cgtcgagatc cgttcactaa tcgaatggat ctgtctctgt ctctctctcc
     5521 accttcttct tctattcctt cgggcctgtc gggtcccctc ggggttggga ggtgggtctg
     5581 aaacgataat ggtgaatatc cctgcctaac tctattcact atagaaagta cagcaaaaac
     5641 tattcttaaa cctaccaagc ctcctactat cattatgaat aattttatat accacagcca
     5701 atttgttatg ttaaaccaat tccacaaact tgcccattta tctaattcca ataattcttg
     5761 ttcattcttt tcttgctggt tttgcgattc ttcaattaag gagtgtatta agcttgtgta
     5821 attgttaatt tctctgtccc actccatcca ggtcgtgtga ttccaaatct gttccagaga
     5881 tttattactc caactagcat tccaaggcac agcagtggtg caaatgagtt ttccagagca
     5941 accccaaatc cccaggagct gttgatcctt taggtatctt tccacagcca ggattcttgc
     6001 ctggagctgc ttgatgcccc agactgtgag ttgcaacaga tgctgttgcg cctcaatagc
     6061 cctcagcaaa ttgttctgct gctgcactat accagacaat aattgtctgg cctgtaccgt
     6121 cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct cccaagaacc caaggaacaa
     6181 agctcctatt cccactgctc ttttttctct ctgcaccact cttctctttg ccttggtggg
     6241 tgctactcct aatggttcaa tttttactac tttatattta tataattcac ttctccaatt
     6301 gtccctcata tctcctcctc caggtctgaa gatcagcggc cgcttgctgt gcggtggtct
     6361 tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta
     6421 tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg
     6481 atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt
     6541 ctaacaggcc aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt
     6601 ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa
     6661 ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag
     6721 tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag
     6781 caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt
     6841 tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     6901 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     6961 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     7021 ccagagagac ccgtttatgt atcgagctag gcacttaaat acaatatctc tgcaatgcgg
     7081 caattcagtg gttcgtccaa tccatgtcag acccgtctgt tgccttccta ataaggcacg
     7141 atcgtaccac cttacttcca ccaatcggca tgcacggtgc tttttctctc cttgtaaggc
     7201 atgttgctaa ctcatcgtta ccatgttgca agactacaag agtattgcat aagactacat
     7261 taagcttgca gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     7321 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7381 cgagccggaa gcataaagtg gaaagcctgg ggt
//
