LOCUS       sequence-192972-        8070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral empty backbone, EGFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8070)
  TITLE     Perkins lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8070)
  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            "Unpublished"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: pKAM-GFP
FEATURES             Location/Qualifiers
     source          1..8070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2221..2432
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2433..3371
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3328..3348
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(3464..3487)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3847..3865
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      3944..3953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3950..3952,3953..3955,3956..4669)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3950..3952,3953..3955,3956..4666)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3995..4016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4256..4275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4603..4624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4788..5421
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5550..5566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5550..5566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5563..5585)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5574..5590
                     /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(5598..5604,5605..5622,5623..5628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5643..5664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5781..5798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5952..6537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6032..6051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6708..7499,7500..7568))
                     /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     7331..7350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7569..7673)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7721..7855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7758..7777)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7812..7831
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt
     2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3361 ttccatttca ggtgtcgtga ggatccgcgg ccgcgccggc tctagatcgc gaacgcgtga
     3421 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     3481 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     3541 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     3601 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     3661 agtagcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg
     3721 ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg
     3781 ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc
     3841 cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt
     3901 tgaccgaatc accgacctct ctccccaggg ggatccaccg gtcgccacca tggtgagcaa
     3961 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     4021 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     4081 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4141 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4261 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4321 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4381 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4441 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4561 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4621 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcgt
     4681 cgacgggccc tcgacgggcc cgcggaattc gagctcggta cctttaagac caatgactta
     4741 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat
     4801 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     4861 ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg
     4921 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa
     4981 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt
     5041 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga
     5101 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc
     5161 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc
     5221 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5281 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5341 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5401 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5461 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg
     5521 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5581 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5641 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5701 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5761 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5821 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5881 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5941 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6001 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6061 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6121 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6181 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6241 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6301 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6361 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6421 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6481 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6541 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6601 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6661 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6721 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6781 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6841 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6901 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6961 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7021 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7081 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7141 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7201 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7261 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7321 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7381 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7441 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7501 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7561 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7621 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7681 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
     7741 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7801 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
     7861 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
     7921 caattaccta gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     7981 aaagaaattg tatttgttaa atatgtacta caaacttagt agtttttaaa gaaattgtat
     8041 ttgttaaata tgtactacaa acttagtagt
//
