LOCUS       sequence-404918-        7514 bp    DNA     circular SYN 15-APR-2025
DEFINITION  transfer plasmid for lentiviral vector production with miR for Hs 
            ATP10B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Wouters R, Beletchi I, Van den Haute C, Baekelandt V, Martin S, 
            Eggermont J, Vangheluwe P
  TITLE     The lipid flippase ATP10B enables cellular lipid uptake under stress
            conditions.
  JOURNAL   Biochim Biophys Acta Mol Cell Res. 2024 Feb;1871(2):119652. doi: 
            10.1016/j.bbamcr.2023.119652. Epub 2023 Dec 11.
  PUBMED    38086447
REFERENCE   2  (bases 1 to 7514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochim 
            Biophys Acta Mol Cell Res."; date: "2024-02"; volume: 
            "1871(2):119652. doi"; pages: " 10.1016/j.bbamcr.2023.119652. Epub 
            2023 Dec 11"
COMMENT     Sequence Label: miR5 HsATP10B
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        754..1133
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1134..1337
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1288..1308
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3027..3434
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             3458..3856
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     ncRNA           3921..3935
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    4028..4616
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4081..4101)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4499..4510)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5324..5342)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5363..5379)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5363..5379)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5376..5398)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5387..5403
                     /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(5411..5417,5418..5435,5436..5441))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5456..5477
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5594..5611)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5765..6353)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5845..5864)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6524..7315,7316..7384))
                     /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     7147..7166
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7385..7489)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctccaccgcg
      661 gtggcggccg ctctagaatt ccattgcata cgttgtatcc atatcataat atgtacattt
      721 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
      781 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      841 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      901 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      961 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     1021 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     1081 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     1141 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     1201 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     1261 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     1321 tctatataag cagagctcgt ttagtgaacc gcagtcgctc tgcggagagg ctggcagatt
     1381 gagccctggg aggttctctc cagcactagc aggtagagcc tgggtgttcc ctgctagact
     1441 ctcaccagca cttggccagt gctgggcaga gtggctccac gcttgcttgc ttaaagacct
     1501 cttcaataaa gctgccattt tagaagtaag ccagtgtgtg ttcccatctc tcctagtcgc
     1561 cgcctggtca actcggtact cggtaataag aagaccctgg tctgttagga ccctttctgc
     1621 tttgagaaac cgaagcagga aaatccctag cagattggcg cccgaacagg gacttgaagg
     1681 agagtgagag actcctgagt acggctgagt gaaggcagta agggcggcag gaaccaacca
     1741 cgacggagtg ctcctataaa ggcgcgggtc ggtaccagac ggcgtgagga gcgggagagg
     1801 aggaggcctc cggttgcagg taagtgcaac acaaaaaaga aatagctgtc ttgttatcca
     1861 ggaagggata ataagataga gtgggagatg ggcgcgagaa actccgtctt gtcagggaag
     1921 aaagcagatg aattgaaaaa attattaatc gcatgaattt taaaagaagg ggaggaatag
     1981 gggatatgac tccagcagaa agattaatta acatgatcac tacagaacaa gaaatacaat
     2041 ttcaacaatc aaaaaactca aaatttaaaa attttcgggt ctattacaga gctcacgcgt
     2101 gattggagtt gggagattat aaattagtag agatcactcc gattggcttg gcccccacag
     2161 atgtgaagag gtacactact ggtggcacct caagaaataa aagaggggtc tttgtgctag
     2221 ggttcttggg ttttctcgca acggcaggtt ctgcaatggg cgcggcgtcg ttgacgctga
     2281 ccgctcagtc ccggacttta ttggctggga tagtgcagca acagcaacag ctgttggacg
     2341 tggtcaagag acaacaagaa ttgttgcgac tgaccgtctg gggaacaaag aacctccaga
     2401 ctagggtcac tgccatcgag aagtacttaa aggaccaggc gcagctaaat gcttggggat
     2461 gtgcgtttag acaagtctgc cacactactg taccatggcc aaatgcaagt ctaacaccag
     2521 actggaacaa tgatacttgg caagagtggg agcgaaaggt tgacttcttg gaggaaaata
     2581 taacagccct cctagaagag gcacaaattc aacaagagaa gaacatgtat gaattacaaa
     2641 agttgaatag ctgggatgtg tttggcaatt ggtttgacct tgcttcttgg ataaagtata
     2701 tacaatatgg aatttatgta gttgtaggag taatactgtt aagaatagtg atctatatag
     2761 tacaaatgct agctaagtta aggcaggggt ataggccagt gttctcttcc ccaccctctt
     2821 atttccagta gactcatacc caacaggacc cggcactgcc aaccagagaa ggcaaagaag
     2881 gacggtggag aaggcggtgg atctgctagc actagttatt aatcgtacga ttaactgcag
     2941 ccccgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3001 cctgtaggtt tggcaagcta gctgcagtaa cgccattttg caaggcatgg aaaaatacca
     3061 aaccaagaat agagaagttc agatcaaggg cgggtacatg aaaatagcta acgttgggcc
     3121 aaacaggata tctgcggtga gcagtttcgg ccccggcccg gggccaagaa cagatggtca
     3181 ccgcagtttc ggccccggcc cgaggccaag aacagatggt ccccagatat ggcccaaccc
     3241 tcagcagttt cttaagaccc atcagatgtt tccaggctcc cccaaggacc tgaaatgacc
     3301 ctgcgcctta tttgaattaa ccaatcagcc tgcttctcgc ttctgttcgc gcgcttctgc
     3361 ttcccgagct ctataaaaga gctcacaacc cctcactcgg cgcgccagtc ctccgacaga
     3421 ctgagtcgcc cgggggaccg ggagcagatc taccggtatg gccaagcctt tgtctcaaga
     3481 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
     3541 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
     3601 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
     3661 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
     3721 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
     3781 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
     3841 tgtgtgggag ggctaactcg agatccaaga aggtatattg ctgttgacag tgagcgtcct
     3901 taagctagta cctatatata gtgaagccac agatgtatat ataggtacta gcttaaggtt
     3961 gcctactgcc tcggacttca agggctacga tgtcgacttg gatccatcga tggatctaat
     4021 tcccgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4081 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4141 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4201 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4261 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4321 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4381 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4441 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4501 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4561 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     4621 tcgagtttta taaaagaaaa ggggggactg gaagggattt attacagtgc aagaagacat
     4681 agaatcttag acatgtactt agaaaaggaa gaaggcatca taccagattg gcaggattac
     4741 acctcaggac caggaattag atacccaaag acatttggct ggctatggaa attagtccct
     4801 gtaaatgtat cagatgaggc acaggaggat gagaggcatt atttaatgca gccagctcaa
     4861 acttccaagt gggatgaccc ttggggagag gttctagcgt ggaagtttga tccaactcta
     4921 gcctacactt atgaggcata tgctagatac ccagaagagt tggaagcaag tcaggcctgt
     4981 cagaactgca tttcgctctg tattcagtcg ctctgcggag aggctggcag attgagccct
     5041 gggaggttct ctccagcact agcaggtaga gcctgggtgt tccctgctag actctcacca
     5101 gcacttggcc agtgctgggc agagtggctc cacgcttgct tgcttaaaga cctcttcaat
     5161 aaagctgcca ttttagaagt aagccagtgt gtgttcccat ctctcctagt cgccgcctgg
     5221 tcaactcggt actcggtaat aagaagaccc tggtctgtta ggaccctttc tgctttgaga
     5281 aaccgaagca ggaaaatccc tagcatggta ccagcttttg ttccctttag tgagggttaa
     5341 tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5701 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5761 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5821 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5881 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5941 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6001 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6061 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6121 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6181 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6241 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6301 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6361 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6421 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6481 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6541 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6601 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6661 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6721 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6781 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6841 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6901 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6961 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7021 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7081 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7141 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7201 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7261 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7321 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7381 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7441 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7501 gaaaagtgcc acct
//
