LOCUS       sequence-237551-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of hFerritinL in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5864)
  TITLE     Trevor Douglas lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5864)
  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: 
            "Unpublished"
COMMENT     Sequence Label: hFerritinL
FEATURES             Location/Qualifiers
     source          1..5864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     172..191
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        172..190
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    191..215
                     /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             230..252
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             896..913
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(966..984)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      980..1027
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1064..1519
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1151..1170)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1361..1382
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1611..2426)
                     /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             complement(1611..2426)
                     /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     2334..2353
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2418..2437
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2548..3136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3037..3056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3290..3307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3322..3461
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3387..3406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3547..3569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3563..3754)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4563..5645)
                     /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     5607..5626
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5646..5723)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
       61 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
      121 ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac
      181 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
      241 ttaagaagga gatatacata tgagctccca gattcgtcag accgttattc cctcactctc
      301 aagcacgact aaggatccgg tatgagctcc cagattcgtc agaattattc caccgacgtg
      361 gaggcagccg tcaacagcct ggtcaatttg tacctgcagg cctcctacac ctacctctct
      421 ctgggcttct atttcgaccg cgatgatgtg gctctggaag gcgtgagcca cttcttccgc
      481 gaactggccg aggagaagcg cgagggctac gagcgtctcc tgaagatgca aaaccagcgt
      541 ggcggccgcg ctctcttcca ggacatcaag aagccagctg aagatgagtg gggtaaaacc
      601 ccagacgcca tgaaagctgc catggccctg gagaaaaagc tgaaccaggc ccttttggat
      661 cttcatgccc tgggttctgc ccgcacggac ccccatctct gtgacttcct ggagactcac
      721 ttcctagatg aggaagtgaa gcttatcaag aagatgggtg accacctgac caacctccac
      781 aggctgggtg gcccggaggc tgggctgggc gagtatctct tcgaaaggct cactctcaag
      841 cacgactaag gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca
      901 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
      961 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1021 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1081 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1141 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1201 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1261 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1321 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1381 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1441 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1501 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1561 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1621 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1681 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1741 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1801 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1861 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     1921 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     1981 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2041 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2101 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2161 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     2221 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2281 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2341 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     2401 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2461 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3301 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3361 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3421 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3481 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3541 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3601 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3661 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3721 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3781 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3841 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     3901 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     3961 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     4021 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4081 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4141 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     4201 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     4261 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     4321 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     4381 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4441 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4501 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     4561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4621 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     4681 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     4741 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     4801 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     4861 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     4921 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     4981 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5041 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5101 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     5161 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     5221 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     5281 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     5341 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     5401 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     5461 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     5521 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     5581 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     5641 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     5701 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     5761 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     5821 atgcaaggag atggcgccca acagtccccc ggccacgggg cctg
//
