LOCUS       sequence-464149-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heterologous protein expression of oxysterol-binding protein homolog
            1 in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  TITLE     AI Proteins, Inc. plasmids
REFERENCE   2  (bases 1 to 6386)
  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: pAIP4_5H28
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     167..186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(310..357)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     353..371
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(424..441)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1555..1584)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1607..1629
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1644..1668
                     /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     complement(1668..1687)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1669..1687)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1775..1794)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2000..2077
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2078..3160
                     /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     complement(2097..2116)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3969..4160
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4154..4176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4262..4401
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4317..4336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4416..4433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4667..4686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5286..5305)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5297..6112
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5297..6112
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5370..5389)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(join(6204..6386,1..273))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6341..6362)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
       61 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
      121 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
      181 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      241 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc
      301 tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta
      361 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
      421 tcagtggtgg tggtggtggt gctcgagtca ttacgaaccg gtcacgtcga tgacagactg
      481 ttcacgggtt gcgcgctcca ggagtttctt cagctcgata tcaatggcga ttttcgtatt
      541 ggtcgctgtg tcgttctcgt tcaccttgcg aactaactca atcggcagtt tacctttacg
      601 atcgcgttta aacggatccg cgccgtgctc caacagccag cggcacatga taacgtcgcg
      661 cttcttcacg aattcgtgca gaacggtatc gccggtttcc ggatccatgc cgttgatgtc
      721 caacaattca gcattacgcg gcgagctcag aatggactcc aaatggccga agtcacggtt
      781 gttgaacgcg gtacgaaact cttgcgccgt ttccgcaaca taggtgctac gcttcacctg
      841 catcatctgg gcaatgttca ggttcttgca catctcaatt gcctgcagat gggagttgtt
      901 cagaacacaa tcgttaatgg tcggctgatc taataggaac gccacgatgt caccacgaga
      961 ctggtatgcc gcaatgtgca gcggagtatt accattgctg tctctttcgt tcaagtccaa
     1021 atggatattc agaaaagtgg tgttcgtggt gctaacccag tgatgcacaa tttctttgat
     1081 gaccgccatc ggagccactt gtacagcgta gtgcagcata aggtgaagga cctgttgaac
     1141 atttggatcg tccagcggtt ggaattgctt tttcagcaga tcttgcagat tcggaaagct
     1201 accctgacgc agcgcgtcta gcaacttaag ttttaacagg ggttttgatc cagagccgcc
     1261 aatctgttca cggtgggctt caatgatgtc gttgtcttcc atgtccaggt cttccggcgc
     1321 ttgatctgcc tggatgcgaa taccgtcata caggaaacgc aggctatcca tttccttgcc
     1381 ctgacgtttg gcaaacgctt ccatcaggcg acgcagcggc gtggtctttt tgatcttaaa
     1441 gaaaatttca gagctgccgt cactaacttt caggttaata tgggtttccg gtttcacttc
     1501 cggcttgact tccggctttg cttcttggtt gacttccgaa tcttgcaggc taccgtggtg
     1561 atggtgatgg tgatgatggt gatgaatctt cgccatcata tgtatatctc cttcttaaag
     1621 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
     1681 cgtattaatt tcgcgggatc gagatcgatc tcgatcctct acgccggacg catcgtggcc
     1741 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     1801 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     1861 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     1921 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     1981 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     2041 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     2101 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     2161 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     2221 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     2281 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     2341 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     2401 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     2461 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     2521 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     2581 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     2641 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     2701 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     2761 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     2821 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggacat
     2881 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     2941 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     3001 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     3061 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3121 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     3181 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
     3241 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     3301 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     3361 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     3421 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     3481 ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc
     3541 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
     3601 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
     3661 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
     3721 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
     3781 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
     3841 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
     3901 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
     3961 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
     4021 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
     4081 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
     4141 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     4201 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     4261 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     4321 atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca cagatgcgta
     4381 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4441 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4501 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag
     5281 taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt
     5341 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     5401 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     5461 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     5521 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     5581 gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga
     5641 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     5701 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     5761 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     5821 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     5881 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     5941 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     6001 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     6061 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6181 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     6241 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6301 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6361 actattaaag aacgtggact ccaacg
//
