LOCUS       sequence-430154-        5492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shanbhag lab plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5492)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 5492)
  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: Stag-ObMT
FEATURES             Location/Qualifiers
     source          1..5492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(11..599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(91..110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(710..729)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             721..1536
                     /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             721..1536
                     /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(794..813)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(1628..2083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1765..1786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1977..1996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2120..2167)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2163..2181
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2234..2251)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2435..2452)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2468..2512)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     RBS             2523..2545
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2560..2584
                     /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(2584..2603)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2585..2603)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2691..2710)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2916..2993
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2994..4076
                     /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(3013..3032)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4885..5076
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5070..5092)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5178..5317
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5233..5252
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5332..5349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
       61 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      421 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
      481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      661 ggattttggt catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
      721 atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat
      781 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
      841 tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
      901 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
      961 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1021 atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1081 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1141 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     1201 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1261 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     1321 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1381 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1441 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1501 ttgcagtttc atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat
     1561 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     1621 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1681 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1741 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1801 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     1861 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     1921 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     1981 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2041 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg
     2101 atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     2161 atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag
     2221 cagccggatc tcagtggtgg tggtggtggt gctcgagtta ttatttgtgg cactcgcagc
     2281 cggtgtgacc gcaaccggtg ccgttcggat gaccatccgc gcacgcgctg ctgcaatagc
     2341 ttttgccgct cttttcaatc gcctcggtca ggctaaccac gcacaggcag ctcgggcacg
     2401 cgcatttgat ctgggtcacg gtggtcatgc ccatggaacc gcgtggcacc agggtaccca
     2461 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta
     2521 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
     2581 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
     2641 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     2701 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     2761 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     2821 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     2881 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
     2941 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     3001 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     3061 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     3121 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     3181 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     3241 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     3301 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     3361 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     3421 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     3481 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     3541 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     3601 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     3661 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     3721 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     3781 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     3841 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     3901 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     3961 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4021 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4081 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg
     4141 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca
     4201 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc
     4261 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta
     4321 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc
     4381 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc
     4441 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
     4501 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
     4561 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
     4621 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
     4681 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
     4741 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
     4801 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
     4861 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     4921 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     4981 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     5041 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5101 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5161 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     5221 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat
     5281 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     5341 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5401 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5461 gcaaaaggcc aggaaccgta aaaaggccgc gt
//
