LOCUS       sequence-460507-        8357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construction of extracellular magnetite accumulating bacterial 
            systems..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8357)
  AUTHORS   Yavuz M, Utkur M, Kehribar ES, Yagiz E, Saritas EU, Seker UOS
  TITLE     Engineered Bacteria with Genetic Circuits Accumulating Nanomagnets 
            as MRI Contrast Agents.
  JOURNAL   Small. 2022 Jul;18(26):e2200537. doi: 10.1002/smll.202200537. Epub 
            2022 May 13.
  PUBMED    35567331
REFERENCE   2  (bases 1 to 8357)
  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: "Small."; 
            date: "2022-07"; volume: "18(26):e2200537. doi"; pages: " 
            10.1002/smll.202200537. Epub 2022 May 13"
COMMENT     Sequence Label: ECMAB - pET22b Ag43 160N 6H Mms6
FEATURES             Location/Qualifiers
     source          1..8357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..147
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     63..82
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(162..179)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(333..921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(413..432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1092..1883,1884..1952))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1715..1734
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1953..2057)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2083..2538)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2220..2241)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2432..2451
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2575..2622)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2618..2636
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2689..2706)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5617..5634)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     sig_peptide     complement(5641..5706)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     RBS             5714..5736
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5751..5775
                     /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(5775..5794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5776..5794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5878..5897)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6103..6180
                     /gene="lacI"
                     /label=lacI promoter
     CDS             6181..7263
                     /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(6200..6219)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             8072..8263
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(8257..8279)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
       61 tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat accgcacaga
      121 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
      181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
      781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1381 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     1441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     1921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     1981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     2101 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2161 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2221 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2281 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2341 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2401 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2461 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2521 cgccgctaca gggcgcgtcc cattcgccaa tccggatata gttcctcctt tcagcaaaaa
     2581 acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc
     2641 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg
     2701 gtggtgctcg agtcatcaga aggtcacatt cagtgtggcc tgaccgttat acccttcagc
     2761 gctgctgccg ctgacgctgt gggcataacc ggcctgaacg cccagggtga tattttcccg
     2821 gacacgggct tccagtccgg cctgcaggtc cagtgatgtg ccattctgtg agggagagaa
     2881 cgtcatcccg ctgcctgcag tggaagtccc cacacgcata tctccccggg agctgaaggt
     2941 gcggataaca gaaggctgta cccaccagtt caccggtaat tcactcacac tgtgttttgc
     3001 actgtcacgc aggggggcac gggatgaggt gccttcgcca aaggtcatat cgttgtggct
     3061 gcccagacgg aaaccggcac gcacatgttg tgcactgcca tgcccgaact tcacataacc
     3121 ggcgttgtcc ttaccgtcat ccagggaaag tccctgccag gtatactgca gttgtggctc
     3181 cagcatcagg ttgtcagtga tactgaaggg cagaccggtt tccagtgagc ccagccagcc
     3241 ccagccccgg gcgcggaagt cgttattgtc cgatgacgct ttcatgctgt ggcgggttcc
     3301 ctgtgccaca atgtcagccc acaggccgga ggacgtgtgt accagattca ggtatccgcc
     3361 caggctgccg gcatcatccc ggaccgtgcc ggcacgggag ccgtcatcat ccttaacatc
     3421 aacggaagaa tggccagcag caccatatac ccccgcggtc acagacatac cggcaacctc
     3481 tgttctcatc aggtcaccct ccagacggac gaatccatag ctgccgctgc tttccggcgt
     3541 ggccccacgg gcaataccgc cattgttatc gtgaccgaga tgaccgccct gaatgctgag
     3601 acggacgctg ttgttttcac catttacacc ggtctgatgg ctgcgggagc ctgccacaat
     3661 ccggtcatag tccattgcct gtgtcagcat ggaggcatac agggggactt ctgcacgata
     3721 agcattttca ctgcgcagat accagctctc atcactgtcc cggttgaggg agtagttaaa
     3781 ggcaccggcc tgcagcctgt tcccctggac aaaggcccct tcctccgtgg tggcaccgtt
     3841 aatggcttcc accacctgaa tacccttacc gctggtcgcc agccccgacg cactgttgcc
     3901 ggcgttcacc aggttcagga tggtttttcc ggttgccctg ccgccgtcaa tgaccagtct
     3961 gtcagcattg ttctgtgcca tatccgggcg tacacgcagg ctgatggtgc cattctgtcc
     4021 gttcaggttt ttcactttca gggttgccgg tacgaacttc cctgtgcggg tggaggtgaa
     4081 atgaatctgt ccggcatggc tgaggtcatc caccaccgac tgcaccgtgg cgttatcggg
     4141 gatattccag gtggcaccgg aggcgagagt gacattcgtg gggtcaatgg caccgttcag
     4201 cacagtgctg ccggtcagct tcagggctgt accgcgctga gcaatgacat ccgtggtgac
     4261 ggtactgtcg ttcagcgtca gcgtgccttc attcaggttg acggctttct gggtgagtgt
     4321 ggtgttgctg acagtgagtg tgccacttcc tgatttttcc acgctgccgt taccggtgag
     4381 agagcctccc tgcaggagtg catcgccttc acggatggtc agggtatcgt tgttcaccgt
     4441 cttcccggaa agggtaagta tggcgccgtt attcagcgtg gtggtgcccg ccagtgtgcc
     4501 gcccctggcg gtgaacagtc cgccatttac cgtggtatcc gtggccgtat caccggcgtt
     4561 cagcgtgaat gaactgcctt tttccagcat cagggcatcc gcctgaccgc ctccgatact
     4621 gaatgccttt ccgtcgctcc gggtaccact gacagcggca ccggaatcgg ccagcagtac
     4681 accgccattt cccatggata cggtggtggc ggtgccaccg ttgcggacat ccagcgttcc
     4741 gccatcatcc acgcgggtat tagtggctgt gtgtccggtc agcacatcca ggcgtccgcc
     4801 attttccagt acgacattat cagctttacc ctccacaaca gagaatgctc ccaggcggtt
     4861 tatgccggta acggttgcag ccgtactggt aaccagtgcg ccgccctgct tcagggtgac
     4921 attcgtggct gtaccaccgg cgtccacctg cagtctgccc ttctgattaa cggtggtatt
     4981 cccggccaca cccccgtttt tgacaatctg ccagccgtca ctgttcacaa cagtgtcaga
     5041 cgctgtaccg ccgttgtgca catactggta tcccccattc agcgtggtat ccagtgcgtg
     5101 accatgtaca gtctggtcgc cgccggcata aaccacagtg gtatttgccg ttccttcagc
     5161 tctcacaatc tggcgaccgt ttttattgat ggttgtgcgc ttaagtgcaa gtgcgtcacg
     5221 aagctcgact tcctcgtctg actgtgcaga ttcaatatca cgactcttca tgtaggcgta
     5281 aaccgctcca gcccctacga cacccaaaat aatcggtccc cacgcaccaa ggcctaagcc
     5341 aagacccagt cctaaccctt tacctgtcca gatggtccct ccaactactt tggcgccagc
     5401 agcggccttt cctgcaccta aggctacctt agtgccagca ccctgagccg cagccacgtt
     5461 agctggtgcg gccggtgctg ccaccgcggt cgcaacgccg gtcttggccg caattccagt
     5521 tttcgctaca gttccggtct tggcagctcc tgttttagcg gcgcctgcct tagcagccgc
     5581 cccttcgcgc tccatttcgc cggatccacc gccaccgtgg tggtggtggt ggtggcccat
     5641 ggccatcgcc ggctgggcag cgaggagcag cagaccagca gcagcggtcg gcagcaggta
     5701 tttcatatgt atatctcctt cttaaagtta aacaaaatta tttctatagg ggaattgtta
     5761 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc
     5821 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct
     5881 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt
     5941 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct
     6001 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct
     6061 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     6121 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     6181 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     6241 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     6301 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     6361 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     6421 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     6481 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     6541 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc
     6601 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt
     6661 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag
     6721 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
     6781 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
     6841 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
     6901 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
     6961 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca
     7021 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
     7081 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
     7141 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
     7201 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     7261 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
     7321 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     7381 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     7441 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct
     7501 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa
     7561 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat
     7621 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     7681 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     7741 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     7801 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     7861 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     7921 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     7981 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     8041 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     8101 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     8161 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     8221 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     8281 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     8341 agcgggtgtt ggcgggt
//
