LOCUS       sequence-180618-        7425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of mTagBFP2 using PGK promoter in a 
            lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7425)
  TITLE     Torok-Storb lab lentiviral fluorescent protein
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7425)
  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            "Unpublished"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: pRRlsinPGK_BFP2_WPRE
FEATURES             Location/Qualifiers
     source          1..7425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..33)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(187..775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(267..286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(946..1737,1738..1806))
                     /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     1569..1588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1807..1911)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1937..2392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2074..2095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2286..2305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2519..2541
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2533..2550
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2534..2550
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2560..2579
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2560..2578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2599..2734
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2774..2895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2785..2804)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2839..2858
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2967..3200
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3287..3875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3393..3404
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3802..3822
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3937..4647)
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      4644..4653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4669..5179)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4732..4750)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_feature    5225..5342
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5609..5653)
                     /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    5838..6071
                     /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    6564..6689
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6736..6916
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6917..7143)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(7171..7191)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7171..7189)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7210..7226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7210..7226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7223..7245)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7234..7250
                     /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(join(7258..7264,7265..7282,7283..7288))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
       61 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      361 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      601 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
      961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     1921 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1981 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2041 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2101 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2161 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2221 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2281 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2341 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2401 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2461 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     2521 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     2581 gaattgggta cgtccctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     2641 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     2701 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggatagcta
     2761 gagccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2821 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2881 tgcaataaac aagttcctct cactctctga tattcatttc tttgcaagtt ataaatactg
     2941 aataataaga tgacatgaac tactactgct agagattttc cacactgact aaaagggtct
     3001 gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt gaagcactca
     3061 aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga gctcccaggc
     3121 tcagatctgg tctaaccaga gagacccagt acaagcaaaa agcagatctt gtcttcgttg
     3181 ggagtgaatt agcccttcca gtcccccctt ttcttttaaa aagtggctaa gatctacagc
     3241 tgccttgtaa gtcattggtc ttaaaggtac cgagctcgaa ttccaggcgg ggaggcggcc
     3301 caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc
     3361 ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag
     3421 gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccatggaaa ggacgtcagc
     3481 ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg
     3541 gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg
     3601 gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca
     3661 gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac
     3721 caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa
     3781 ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc
     3841 aatctttcac aaattttgta atccagaggt tgattgtcga ctgcagttcg aacccgggca
     3901 tatgtctaga accggtggat cagtcgcggc cgcttaatta agcttgtgcc ccagtttgct
     3961 agggaggtcg cagtatctgg ccactgccac ctcgtgctgc tcgacgtagg tctcgttgtt
     4021 ggcctccttg attctttcca gtctgtagtc cacatagtag acgccaggca tcttgaggtt
     4081 cttagcgggt ttcttggatc tatatgtggt cttggcgttt gcgatcagat ggctcccgcc
     4141 cacgagcttc agggccatgt cgtttctgcc ttccaggccg ccgtcagcgg ggtacagcgt
     4201 ctcggtgaag gcctcccagc cgagtgtttt cttctgcatc acagggccgt tggatgtgaa
     4261 gttcacccct ctgatcttga cgttgtagat gaggcagccg tcctggaggc tggtgtcctg
     4321 ggtagcggtc agcacgcccc cgtcttcgta tgtggtgact ctctcccatg tgaagccctc
     4381 agggaaggac tgcttgaaga agtcggggat gccctgggtg tggttgatga aggtcttgct
     4441 gccgtagagg aagctagtag ccaggatgtc gaaggcgaag gggagagggc cgccctcgac
     4501 caccttgatt ctcatggtct gggtgccctc gtagggcttg ccttcgccct cggatgtgca
     4561 cttgaagtga tggttgtcca cggtgccctc catgtacagc ttcatgtgca tgttctcctt
     4621 aatcagctct tcgcccttag acaccatggt ggcgaccggt ggatccccct ggggagagag
     4681 gtcggtgatt cggtcaacga gggagccgac tgccgacgtg cgctccggag gcttgcagaa
     4741 tgcggaacac cgcgcgggca ggaacagggc ccacactacc gccccacacc ccgcctcccg
     4801 caccgcccct tcccggccgc tgctctcggc gcgccctgct gagcagccgc tattggccac
     4861 agcccatcgc ggtcggcgcg ctgccattgc tccctggcgc tgtccgtctg cgagggtact
     4921 agtgagacgt gcggcttccg tttgtcacgt ccggcacgcc gcgaaccgca aggaaccttc
     4981 ccgacttagg ggcggagcag gaagcgtcgc cggggggccc acaagggtag cggcgaagat
     5041 ccgggtgacg ctgcgaacgg acgtgaagaa tgtgcgagac ccagggtcgg cgccgctgcg
     5101 tttcccggaa ccacgcccag agcagccgcg tccctgcgca aacccagggc tgccttggaa
     5161 aaggcgcaac cccaaccccg tggaattcga tatcaagctt ctcgaggcta gtctcgtgat
     5221 cgataaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt atgtctgttg
     5281 ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct tttcttttaa
     5341 aagttaaccg ataccgtcga gatccgttca ctaatcgaat ggatctgtct ctgtctctct
     5401 ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt gggaggtggg
     5461 tctgaaacga taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa
     5521 aaactattct taaacctacc aagcctccta ctatcattat gaataatttt atataccaca
     5581 gccaatttgt tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt
     5641 cttgttcatt cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg
     5701 tgtaattgtt aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca
     5761 gagatttatt actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag
     5821 agcaacccca aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc
     5881 ttgcctggag ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa
     5941 tagccctcag caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta
     6001 ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga
     6061 acaaagctcc tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg
     6121 tgggtgctac tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc
     6181 aattgtccct catatctcct cctccaggtc tgaagatcag cggccgcttg ctgtgcggtg
     6241 gtcttacttt tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc
     6301 tctatccttt gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct
     6361 tctgatcctg tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat
     6421 gtttctaaca ggccaggatt aactgcgaat cgttctagct ccctgcttgc ccatactata
     6481 tgttttaatt tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat
     6541 ctaattctcc cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg
     6601 ctagtcaaaa tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct
     6661 tcagcaagcc gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc
     6721 ctgttcgggc gccactgcta gagattttcc acactgacta aaagggtctg agggatctct
     6781 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     6841 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     6901 ctaaccagag agacccgttt atgtatcgag ctaggcactt aaatacaata tctctgcaat
     6961 gcggcaattc agtggttcgt ccaatccatg tcagacccgt ctgttgcctt cctaataagg
     7021 cacgatcgta ccaccttact tccaccaatc ggcatgcacg gtgctttttc tctccttgta
     7081 aggcatgttg ctaactcatc gttaccatgt tgcaagacta caagagtatt gcataagact
     7141 acattaagct tgcagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     7201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     7261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     7321 cttaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     7381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgg
//
