LOCUS       sequence-352639-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of human Igf1-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  TITLE     Gerace lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8307)
  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            "Unpublished"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: pLV CMV IGF1-FLAG
FEATURES             Location/Qualifiers
     source          1..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1575..1691
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1714..2017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2018..2221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2172..2192
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2218..2242
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2391..2414
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2740..3328
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2793..2813)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3211..3222)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3342..3841
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3377..3400)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3774..3793
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3862..4461
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3862..3881)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4358..4378
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4599..4832
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4904..5025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4941..4960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4995..5014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5065..5200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5220..5239)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5221..5239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5249..5266)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5249..5265)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5258..5280)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5407..5862
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5494..5513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5704..5725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5888..5992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5993..6061,6062..6853)
                     /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     complement(6211..6230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7024..7612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7513..7532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7766..7783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7900..7921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7936..7941,7942..7959,7960..7966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7974..7990
                     /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     7979..8001
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7998..8014
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7998..8014
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8033..8053
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8035..8053
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8081..8307
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     1681 ttcaaaattt tatcgataag cttgggagtt ccgcgttaca taacttacgg taaatggccc
     1741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2221 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     2281 aagacaccga ctctagagga tccttgaagg tgaagatgca caccatgtcc tcctcgcatc
     2341 tcttctacct ggcgctgtgc ctgctcacct tcaccagctc tgccacggct gactacaagg
     2401 acgacgatga caagggaccg gagacgctct gcggggctga gctggtggat gctcttcagt
     2461 tcgtgtgtgg agacaggggc ttttatttca acaagcccac agggtatggc tccagcagtc
     2521 ggagggcgcc tcagacaggc atcgtggatg agtgctgctt ccggagctgt gatctaagga
     2581 ggctggagat gtattgcgca cccctcaagc ctgccaagtc agctcgctct gtccgtgccc
     2641 agcgccacac cgacatgccc aagacccaga aggaagtaca tttgaagaac gcaagtagag
     2701 ggagtgcagg aaacaagaac tacaggatgt agggtcgaca atcaacctct ggattacaaa
     2761 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2821 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2881 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2941 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3001 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3061 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3121 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     3181 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3241 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3301 cggatctccc tttgggccgc ctccccgcct ggaattctac cgggtagggg aggcgctttt
     3361 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     3421 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     3481 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     3541 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     3601 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     3661 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     3721 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     3781 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     3841 gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3901 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3961 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4021 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4081 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4141 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4201 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4261 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4321 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4381 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4441 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     4501 gcacgacccc atgcatctcg agggcccggt acctttaaga ccaatgactt acaaggcagc
     4561 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctag ctcactccca
     4621 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     4681 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     4741 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     4801 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     4861 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     4921 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4981 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     5041 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5101 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     5161 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     5221 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     5281 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5341 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5401 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5461 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5521 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5581 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5641 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5701 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5761 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5821 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     5881 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5941 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6001 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6061 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6121 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6181 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6241 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6301 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6361 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     6421 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6481 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6541 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6601 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6661 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6721 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6781 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6841 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6901 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6961 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7021 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7081 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7141 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7201 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7261 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7321 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7381 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7441 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7501 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7561 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7621 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7681 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7741 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7801 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7861 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7921 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7981 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     8041 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     8101 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     8161 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     8221 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     8281 tttaagtgcc tagctcgata cataaac
//
