LOCUS       sequence-460634-        6962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Construction of intracellular magnetite accumulating bacterial 
            systems..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6962)
  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 6962)
  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: ICMAB - pET22b T7 EncFtn Mms6
FEATURES             Location/Qualifiers
     source          1..6962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     149..166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    181..320
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(246..265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     406..428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(422..613)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1422..2504)
                     /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     2466..2485
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2505..2582)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2788..2807
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2891..2910
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2891..2909
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2910..2934
                     /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             2946..2957
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3303..3320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4641..4658
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4711..4729)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4725..4772
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4809..5264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4896..4915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5106..5127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5290..5394
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5424..5526
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5527..6186
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5593..5612)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      6369..6957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6858..6877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
       61 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      181 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
      241 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
      301 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
      361 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
      421 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
      481 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
      541 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
      601 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
      661 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
      721 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
      781 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
      841 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
      901 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
      961 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     1021 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     1081 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     1141 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     1201 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     1261 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
     1321 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
     1381 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
     1441 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1501 tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg
     1561 cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc
     1621 aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta
     1681 tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg
     1741 cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc
     1801 tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt
     1861 tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga
     1921 cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg
     1981 accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg
     2041 ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca
     2101 gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg
     2161 agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc
     2221 accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc
     2281 gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt
     2341 tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc
     2401 cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag
     2461 acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat
     2521 tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg
     2581 tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag
     2641 gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa
     2701 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     2761 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     2821 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat
     2881 cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2941 gaattaaaga ggagaaagga tccatggcgc agtccagcaa tagcacccat gaaccactgg
     3001 aagttctgaa agaagaaact gtcaaccgtc atcgtgcaat tgttagcgtg atggaagaac
     3061 tggaagcagt ggactggtat gaccagcgcg tagacgcatc caccgatccg gaactgaccg
     3121 caattctggc ccacaaccgt gatgaggaaa aagaacacgc agcgatgacc ctggaatggc
     3181 tgcgtcgtaa tgatgcaaaa tgggcagaac acctgcgtac ctacctgttc accgagggcc
     3241 cgattacggc tgctaactcc tctagcgtcg ataaactggc tgcggcgctg gaactcgaga
     3301 tgcaccacca ccaccaccac ggcggtagcg gtggcgaaat ggagcgcgaa ggggcggctg
     3361 ctaaggcagg cgccgctaaa acaggagctg ccaagaccgg aactgtagcg aaaactggaa
     3421 ttgcggccaa gaccggcgtt gcgaccgcgg tggcagcacc ggccgcacca gctaacgtgg
     3481 ctgcggctca gggtgctggc actaaggtag ccttaggtgc aggaaaggcc gctgctggcg
     3541 ccaaagtagt tggagggacc atctggacag gtaaagggtt aggactgggt cttggcttag
     3601 gccttggtgc gtggggaccg attattttgg gtgtcgtagg ggctggagcg gtttacgcct
     3661 acatgaagag tcgtgatatt gaatctgcac agtcagacga ggaagtcgag cttcgtgacg
     3721 cacttgcagg cggcagcggc agcctgagca ccgtgcagac cattagcccg agcaaccatt
     3781 gatgaaagct taaggagggt accatgtttg ttccgtttct cattatgttg cgcgaaggac
     3841 ttgaagccgc gctgattgtc agtttgattg ccagctatct taagcgtacc cagcgaggcc
     3901 gatggattgg tgtgatgtgg attggcgtgt tgcttgccgc tgcgttgtgc ctgggcttgg
     3961 gtatcttcat taacgaaacc accggcgaat ttccgcaaaa agaacaggaa ctgtttgaag
     4021 gtatcgtggc ggtgatcgcc gtggtgatcc ttacctggat ggttttctgg atgcgcaaag
     4081 tgtcgcgcaa cgtcaaagtg caactggaac aggcagtcga tagcgcattg cagcgtggaa
     4141 atcatcatgg ctgggcgctg gtgatgatgg tcttttttgc cgttgcaagg gaagggctgg
     4201 agtcggtctt tttcctgctg gcggcatttc aacaagatgt cgggatctgg ccgccgctgg
     4261 gtgcaatgct cggtcttgct actgccgtgg tgctaggctt cctgctctac tggggcggta
     4321 ttcgcctcaa tcttggtgca ttttttaaat ggaccagcct gtttattctc ttcgtcgccg
     4381 cagggctggc agctggtgcc attcgcgcat ttcatgaagc cggattgtgg aaccactttc
     4441 aggaaatcgc cttcgatatg agtgcggtgc tctcaactca ctcgctgttt ggcacgctga
     4501 tggaagggat ttttggctat caggaagcgc cgagcgtcag cgaagtcgcc gtctggttta
     4561 tttatctcat cccggcgctg gtggcatttg ctctgccacc acgcgcaggg gcgacagcgt
     4621 ctcgctccgc gtaactgcag caccaccacc accaccactg agatccggct gctaacaaag
     4681 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     4741 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     4801 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4861 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4921 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4981 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5041 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5101 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5161 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5221 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5281 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5341 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtgacgtc
     5401 gatatctggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat
     5461 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     5521 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     5581 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     5641 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     5701 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     5761 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     5821 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     5881 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     5941 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     6001 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     6061 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     6121 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     6181 gcgtaatttg atatctaact gtcagaccaa gtttactcat atatacttta gattgattta
     6241 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6301 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6361 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6421 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6481 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6541 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6601 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6661 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6721 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6781 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6841 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6901 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6961 gc
//
