LOCUS       sequence-201395-        5966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for expression of BAF (BANF1) in E.coli.Coding sequence in 
            pET15B..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5966)
  AUTHORS   Lee MS, Craigie R
  TITLE     A previously unidentified host protein protects retroviral DNA from 
            autointegration.
  JOURNAL   Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1528-33.
  PUBMED    9465049
REFERENCE   2  (bases 1 to 5966)
  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: "Proc Natl 
            Acad Sci U S A."; date: "1998-02-17"; volume: "95(4)"; pages: 
            "1528-33"
COMMENT     Sequence Label: pBAF
FEATURES             Location/Qualifiers
     source          1..5966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(467..485)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      481..528
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(703..708,709..725,726..731))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(758..776)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        844..948
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(949..1017,1018..1809)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1167..1186)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1980..2568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2469..2488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2722..2739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2754..2893
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2819..2838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2979..3001
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2995..3186)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4498..5580)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5542..5561
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5581..5658)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5864..5883
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg acaacctccc aaaagcaccg agacttcgtg
      181 gcagagccca tgggggagaa gccagtgggg agcctggctg ggattggtga agtcctgggc
      241 aagaagctgg aggaaagggg ttttgacaag gcctatgttg tccttggcca gtttctggtg
      301 ctaaagaaag atgaagacct cttccgggaa tggctgaaag acacttgtgg cgccaacgcc
      361 aagcagtccc gggactgctt cggatgcctt cgagagtggt gcgacgcctt cttgtaggat
      421 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
      481 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
      541 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
      601 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
      661 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
      721 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
      781 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
      841 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      901 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      961 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1021 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1081 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1141 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     1201 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1261 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1321 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1381 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1441 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     1501 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1561 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1621 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1681 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1741 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1801 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1861 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1921 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1981 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2041 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2101 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2161 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2221 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2281 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2341 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2401 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2461 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2521 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2581 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2641 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2701 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2761 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     2821 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     2881 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2941 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3001 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3061 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3121 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3181 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3241 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3301 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     3361 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     3421 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     3481 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     3541 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     3601 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     3661 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     3721 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     3781 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     3841 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     3901 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     3961 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     4021 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     4081 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     4141 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     4201 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     4261 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     4321 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     4381 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     4441 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     4501 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4561 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     4621 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     4681 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     4741 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     4801 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     4861 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     4921 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     4981 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     5041 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     5101 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     5161 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     5221 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     5281 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     5341 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     5401 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     5461 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     5521 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     5581 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     5641 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     5701 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     5761 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     5821 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     5881 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     5941 ggatcgagat ctcgatcccg cgaaat
//
