LOCUS       sequence-380658-        8772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MTP1 (SLC40A1) protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8772)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8772)
  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"
COMMENT     Sequence Label: pLenti6MFV SLC40A1
FEATURES             Location/Qualifiers
     source          1..8772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        42..345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        346..549
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     500..520
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     546..570
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             641..664
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             683..703
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             707..748
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2508..2549
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2586..2606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2591..2920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2771..2906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2968..3015
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3034..3432
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3519..3752
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3824..3945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3861..3880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3915..3934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3985..4120
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4140..4159)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4141..4159)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4169..4186)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4169..4185)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4178..4200)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4327..4782
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4414..4433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4624..4645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4808..4912
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4913..4981,4982..5773)
                     /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(5131..5150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5944..6532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6433..6452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6686..6703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6820..6841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6856..6861,6862..6879,6880..6886)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6894..6910
                     /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     6899..6921
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6918..6934
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6918..6934
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6953..6973
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6955..6973
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7001..7227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7228..7408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7455..7580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8073..8306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8491..8535
                     /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"
ORIGIN
        1 gtgaacggat ctcgacggta tcgataagct tgggagttcc gcgttacata acttacggta
       61 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      121 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      181 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      241 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      301 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      361 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      421 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      481 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      541 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      601 ctccatagaa gacaccgact ctagaggatc cgccaccatg gattacaagg atgacgacga
      661 taagagcccg ggcggatcgt tggagaatct ttattttcag ggtcaaggta agcctatccc
      721 taaccctctc ctcggtctcg attctacgtc tagactgcag gaattcatga ccagggcggg
      781 agatcacaac cgccagagag gatgctgtgg atccttggcc gactacctga cctctgcaaa
      841 attccttctc taccttggtc attctctctc tacttgggga gatcggatgt ggcactttgc
      901 ggtgtctgtg tttctggtag agctctatgg aaacagcctc cttttgacag cagtctacgg
      961 gctggtggtg gcagggtctg ttctggtcct gggagccatc atcggtgact gggtggacaa
     1021 gaatgctaga cttaaagtgg cccagacctc gctggtggta cagaatgttt cagtcatcct
     1081 gtgtggaatc atcctgatga tggttttctt acataaacat gagcttctga ccatgtacca
     1141 tggatgggtt ctcacttcct gctatatcct gatcatcact attgcaaata ttgcaaattt
     1201 ggccagtact gctactgcaa tcacaatcca aagggattgg attgttgttg ttgcaggaga
     1261 agacagaagc aaactagcaa atatgaatgc cacaatacga aggattgacc agttaaccaa
     1321 catcttagcc cccatggctg ttggccagat tatgacattt ggctccccag tcatcggctg
     1381 tggctttatt tcgggatgga acttggtatc catgtgcgtg gagtacgttc tgctctggaa
     1441 ggtttaccag aaaaccccag ctctagctgt gaaagctggt cttaaagaag aggaaactga
     1501 attgaaacag ctgaatttac acaaagatac tgagccaaaa cccctggagg gaactcatct
     1561 aatgggtgtg aaagactcta acatccatga gcttgaacat gagcaagagc ctacttgtgc
     1621 ctcccagatg gctgagccct tccgtacctt ccgagatgga tgggtctcct actacaacca
     1681 gcctgtgttt ctggctggca tgggtcttgc tttcctttat atgactgtcc tgggctttga
     1741 ctgcatcacc acagggtacg cctacactca gggactgagt ggttccatcc tcagtatttt
     1801 gatgggagca tcagctataa ctggaataat gggaactgta gcttttactt ggctacgtcg
     1861 aaaatgtggt ttggttcgga caggtctgat ctcaggattg gcacagcttt cctgtttgat
     1921 cttgtgtgtg atctctgtat tcatgcctgg aagccccctg gacttgtccg tttctccttt
     1981 tgaagatatc cgatcaaggt tcattcaagg agagtcaatt acacctacca agatacctga
     2041 aattacaact gaaatataca tgtctaatgg gtctaattct gctaatattg tcccggagac
     2101 aagtcctgaa tctgtgccca taatctctgt cagtctgctg tttgcaggcg tcattgctgc
     2161 tagaatcggt ctttggtcct ttgatttaac tgtgacacag ttgctgcaag aaaatgtaat
     2221 tgaatctgaa agaggcatta taaatggtgt acagaactcc atgaactatc ttcttgatct
     2281 tctgcatttc atcatggtca tcctggctcc aaatcctgaa gcttttggct tgctcgtatt
     2341 gatttcagtc tcctttgtgg caatgggcca cattatgtat ttccgatttg cccaaaatac
     2401 tctgggaaac aagctctttg cttgcggtcc tgatgcaaaa gaagttagga aggaaaatca
     2461 agcaaataca tctgttgttt gactcgaggg ggggcccgcg gttcgaaggt aagcctatcc
     2521 ctaaccctct cctcggtctc gattctacgc gtaccggtta gtaatgagtt tggaattaat
     2581 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2641 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2701 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2761 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2821 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2881 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2941 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     3001 atagtataat acgacaaggt gaggaactaa accatggcca agcctttgtc tcaagaagaa
     3061 tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac
     3121 agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat
     3181 cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca
     3241 gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc
     3301 tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag
     3361 gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg
     3421 tgggagggct aagcacaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc
     3481 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     3541 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     3601 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     3661 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     3721 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     3781 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     3841 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3901 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     3961 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4021 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4081 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     4141 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     4201 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4261 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4321 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4381 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4441 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4501 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4561 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4621 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4681 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4741 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     4801 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4861 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4921 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4981 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5041 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5101 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5161 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5221 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5281 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5341 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5401 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5461 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5521 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5581 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5641 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5701 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5761 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5821 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5881 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6721 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6781 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     6841 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     6901 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     6961 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     7021 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     7081 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     7141 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     7201 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     7261 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7321 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7381 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     7441 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     7501 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     7561 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     7621 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     7681 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     7741 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     7801 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     7861 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     7921 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     7981 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     8041 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     8101 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     8161 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     8221 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     8281 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     8341 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     8401 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     8461 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     8521 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     8581 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     8641 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     8701 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     8761 tccattcgat ta
//
