LOCUS       sequence-380754-        8106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NTCP (SLC10A1) protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 8106)
  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 SLC10A1
FEATURES             Location/Qualifiers
     source          1..8106
                     /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             1842..1883
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1920..1940)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1925..2254
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2105..2240
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2302..2349
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2368..2766
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2853..3086
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3195..3214)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3249..3268
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3319..3454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3474..3493)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3475..3493)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3503..3520)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3503..3519)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3512..3534)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3661..4116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3748..3767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3958..3979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4142..4246
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4247..4315,4316..5107)
                     /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(4465..4484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5278..5866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5767..5786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6020..6037
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6154..6175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6190..6195,6196..6213,6214..6220)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6228..6244
                     /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     6233..6255
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6252..6268
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6252..6268
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6287..6307
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6289..6307
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6335..6561
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6562..6742
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6789..6914
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7407..7640
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7825..7869
                     /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 gaattcatgg aggcccacaa
      781 cgcgtctgcc ccattcaact tcaccctgcc acccaacttt ggcaagcgcc ccacagacct
      841 ggcactgagc gtcatcctgg tgttcatgtt gttcttcatc atgctctcgc tgggctgcac
      901 catggagttc agcaagatca aggctcactt atggaagcct aaagggctgg ccatcgccct
      961 ggtggcacag tatggcatca tgcccctcac ggcctttgtg ctgggcaagg tcttccggct
     1021 gaagaacatt gaggcactgg ccatcttggt ctgtggctgc tcacctggag ggaacctgtc
     1081 caatgtcttc agtctggcca tgaaggggga catgaacctc agcattgtga tgaccacctg
     1141 ctccaccttc tgtgcccttg gcatgatgcc tctcctcctg tacatctact ccagggggat
     1201 ctatgatggg gacctgaagg acaaggtgcc ctataaaggc atcgtgatat cactggtcct
     1261 ggttctcatt ccttgcacca tagggatcgt cctcaaatcc aaacggccac aatacatgcg
     1321 ctatgtcatc aagggaggga tgatcatcat tctcttgtgc agtgtggccg tcacagttct
     1381 ctctgccatc aatgtgggga agagcatcat gtttgccatg acaccactct tgattgccac
     1441 ctcctccctg atgcctttta ttggctttct gctgggttat gttctctctg ctctcttctg
     1501 cctcaatgga cggtgcagac gcactgtcag catggagact ggatgccaaa atgtccaact
     1561 ctgttccacc atcctcaatg tggcctttcc acctgaagtc attggaccac ttttcttctt
     1621 tcccctcctc tacatgattt tccagcttgg agaagggctt ctcctcattg ccatattttg
     1681 gtgctatgag aaattcaaga ctcccaagga taaaacaaaa atgatctaca cagctgccac
     1741 aactgaagaa acaattccag gagctctggg aaatggcacc tacaaagggg aggactgctc
     1801 cccttgcaca gcctagctcg agggggggcc cgcggttcga aggtaagcct atccctaacc
     1861 ctctcctcgg tctcgattct acgcgtaccg gttagtaatg agtttggaat taattctgtg
     1921 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1981 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2101 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2161 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2221 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     2281 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     2341 taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc
     2401 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc
     2461 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt
     2521 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc
     2581 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga
     2641 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt
     2701 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag
     2761 ggctaagcac aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     2821 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     2881 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     2941 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     3001 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     3061 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     3121 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     3181 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3241 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     3301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3421 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     3481 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     3541 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     3601 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     3661 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3721 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3781 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3841 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3901 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3961 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4021 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4081 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4141 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4201 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4261 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4321 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4381 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4441 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4501 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4561 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4621 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4681 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4741 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4801 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4861 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4921 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4981 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5041 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5101 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5161 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5221 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5281 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5341 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5401 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5461 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5521 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5581 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5641 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5701 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5761 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5821 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5881 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5941 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6001 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6061 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6121 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6181 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6241 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     6301 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     6361 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     6421 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     6481 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     6541 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     6601 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     6661 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     6721 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     6781 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     6841 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     6901 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     6961 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     7021 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     7081 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     7141 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     7201 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     7261 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     7321 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     7381 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     7441 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     7501 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     7561 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     7621 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     7681 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     7741 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     7801 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     7861 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     7921 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     7981 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     8041 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     8101 cgatta
//
