LOCUS       sequence-380659-        7959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ANT1 (SLC25A4) protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  TITLE     Jun Liu Lab Plasmids 2022
REFERENCE   2  (bases 1 to 7959)
  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 SLC25A4
FEATURES             Location/Qualifiers
     source          1..7959
                     /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             1695..1736
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1773..1793)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1778..2107
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1958..2093
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2155..2202
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2221..2619
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2706..2939
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3048..3067)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3102..3121
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3172..3307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3327..3346)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3328..3346)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3356..3373)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3356..3372)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3365..3387)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3514..3969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3601..3620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3811..3832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3995..4099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4100..4168,4169..4960)
                     /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(4318..4337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5131..5719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5620..5639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5873..5890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6007..6028
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6043..6048,6049..6066,6067..6073)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6081..6097
                     /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     6086..6108
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6105..6121
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6105..6121
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6140..6160
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6142..6160
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6188..6414
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6415..6595
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6642..6767
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7260..7493
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7678..7722
                     /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 gaattcgata tcatgggtga
      781 tcacgcttgg agcttcctaa aggacttcct ggccgggggc gtcgccgctg ccgtctccaa
      841 gaccgcggtc gcccccatcg agagggtcaa actgctgctg caggtccagc atgccagcaa
      901 acagatcagt gctgagaagc agtacaaagg gatcattgat tgtgtggtga gaatccctaa
      961 ggagcagggc ttcctctcct tctggagggg taacctggcc aacgtgatcc gttacttccc
     1021 cacccaagct ctcaacttcg ccttcaagga caagtacaag cagctcttct tagggggtgt
     1081 ggatcggcat aagcagttct ggcgctactt tgctggtaac ctggcgtccg gtggggccgc
     1141 tggggccacc tccctttgct ttgtctaccc gctggacttt gctaggacca ggttggctgc
     1201 tgatgtgggc aagggcgccg cccagcgtga gttccatggt ctgggcgact gtatcatcaa
     1261 gatcttcaag tctgatggcc tgagggggct ctaccagggt ttcaacgtct ctgtccaagg
     1321 catcattatc tatagagctg cctacttcgg agtctatgat actgccaagg ggatgctgcc
     1381 tgaccccaag aacgtgcaca tttttgtgag ctggatgatt gcccagagtg tgacggcagt
     1441 cgcagggctg gtgtcctacc cctttgacac tgttcgtcgt agaatgatga tgcagtccgg
     1501 ccggaaaggg gccgatatta tgtacacggg gacagttgac tgctggagga agattgcaaa
     1561 agacgaagga gccaaggcct tcttcaaagg tgcctggtcc aatgtgctga gaggcatggg
     1621 cggtgctttt gtattggtgt tgtatgatga gatcaaaaaa tatgtctaac tcgagggggg
     1681 gcccgcggtt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
     1741 ccggttagta atgagtttgg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     1801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1861 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1921 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     1981 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2041 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     2101 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     2161 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     2221 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
     2281 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
     2341 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
     2401 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
     2461 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
     2521 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
     2581 cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacaattcga gctcggtacc
     2641 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     2701 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     2761 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     2821 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     2881 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     2941 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     3001 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3121 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     3181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3301 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     3361 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3421 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3481 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     3541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3961 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     6001 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     6061 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     6121 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     6181 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
     6241 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
     6301 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
     6361 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
     6421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     6601 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     6661 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     6721 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     6781 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     6841 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     6901 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     6961 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     7021 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     7081 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     7141 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7201 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     7261 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     7321 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     7381 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     7441 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     7501 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     7561 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     7621 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     7681 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     7741 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     7801 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     7861 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     7921 gaaggtggag agagagacag agacagatcc attcgatta
//
