LOCUS       sequence-57411-e        5472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5472)
  AUTHORS   Lelyveld VS, Brustad E, Arnold FH, Jasanoff A
  TITLE     Metal-substituted protein MRI contrast agents engineered for 
            enhanced relaxivity and ligand sensitivity.
  JOURNAL   J Am Chem Soc. 2011 Feb 2;133(4):649-51. doi: 10.1021/ja107936d.
  PUBMED    21171606
REFERENCE   2  (bases 1 to 5472)
  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"; doi: "10"; journalName: 
            "J Am Chem Soc"; date: "2011-02-2- 2"; volume: "133"; issue: "4"; 
            pages: "649-51"
COMMENT     Sequence Label: pChuA
FEATURES             Location/Qualifiers
     source          1..5472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             2069..2113
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2151..2169)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2165..2212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2442..3257)
                     /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(2442..3257)
                     /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     3165..3184
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3258..3349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3365..4114)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3375..3392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3615..3634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5294..5313
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5333..5410)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5456..5472,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5456..5472,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggctcac gtccgcaatt tacctcgttg cgtttgagtt tattggcctt
      121 agctgtttct gccaccttgc caacgtttgc ttttgctact gaaaccatga ccgttacggc
      181 aacggggaat gcccgtagtt ccttcgaagc gcctatgatg gtcagcgtca tcgacacttc
      241 cgctcctgaa aatcaaacgg ctacttcagc caccgatctg ctgcgtcatg ttcctggaat
      301 tactctggat ggtaccggac gaaccaacgg tcaggatgta aatatgcgtg gctatgatca
      361 tcgcggcgtg ctggttcttg tcgatggtgt tcgtcaggga acggataccg gacacctgaa
      421 tggcactttt ctcgatccgg cgctgatcaa gcgtgttgag attgttcgtg gaccttcagc
      481 attactgtat ggcagtggcg cgctgggtgg agtgatctcc tacgatacgg tcgatgcaaa
      541 agatttattg caggaaggac aaagcagtgg ttttcgtgtc tttggtactg gcggcacggg
      601 ggaccatagc ctgggattag gcgcgagcgc gtttgggcga actgaaaatc tggatggtat
      661 tgtggcctgg tccagtcgcg atcggggtga tttacgccag agcaatggtg aaaccgcgcc
      721 gaatgacgag tccattaata acatgctggc gaaagggacc tggcaaattg attcagccca
      781 gtctctgagc ggtttagtgc gttactacaa caacgacgcg cgtgaaccaa aaaatccgca
      841 gaccgttggg gcttctgaaa gcagcaaccc gatggttgat cgttcaacaa ttcaacgcga
      901 tgcgcagctt tcttataaac tcgccccgca gggcaacgac tggttaaatg cagatgcaaa
      961 aatttattgg tcggaagtcc gtattaatgc gcaaaacacg gggagttccg gcgagtatcg
     1021 tgaacagata acaaaaggag ccaggctgga gaaccgttcc actctctttg ccgacagttt
     1081 cgcttctcac ttactgacat atggcggtga gtattatcgt caggaacaac atccgggcgg
     1141 cgcgacgacg ggcttcccgc aagcaaaaat cgattttagc tccggctggc tacaggatga
     1201 gatcacctta cgcgatctgc cgattaccct gcttggcgga acccgctatg acagttatcg
     1261 cggtagcagt gacggttaca aagatgttga tgccgacaaa tggtcatctc gtgcggggat
     1321 gactatcaat ccgactaact ggctgatgtt atttggctca tatgcccagg cattccgcgc
     1381 cccgacgatg ggcgaaatgt ataacgattc taagcacttc tcgattggtc gcttctatac
     1441 caactattgg gtgccaaacc cgaacttacg tccggaaact aacgaaactc aggagtacgg
     1501 ttttgggctg cgttttgatg acctgatgtt gtccaatgat gctctggaat ttaaagccag
     1561 ctactttgat accaaagcga aggattacat ctccacgacc gtcgatttcg cggcggcgac
     1621 gactatgtcg tataacgtcc cgaacgccaa aatctggggc tgggatgtga tgacgaaata
     1681 taccactgat ctgtttagcc ttgatgtggc ctataaccgt acccgcggca aagacaccga
     1741 taccggcgaa tacatctcca gcattaaccc ggatactgtt accagcactc tgaatattcc
     1801 gatcgctcac agtggcttct ctgttgggtg ggttggtacg tttgccgatc gctcaacaca
     1861 tatcagcagc agttacagca aacaaccagg ctatggcgtg aatgatttct acgtcagtta
     1921 tcaaggacaa caggcgctca aaggtatgac cactactttg gtgttgggta acgctttcga
     1981 caaagagtac tggtcgccgc aaggcatccc acaggatggt cgtaacggaa aaattttcgt
     2041 gagttatcaa tggtaactcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca
     2101 gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca
     2161 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     2221 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2281 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc
     2341 gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2401 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     2461 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2521 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2581 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2641 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2701 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2761 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2821 acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2881 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     2941 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3001 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3061 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3121 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3181 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     3241 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     3301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcag
     3361 cgctcttccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     3421 gtgtcagctc actcaaaagc ggtaatacgg ttatccacag aatcagggga taaagccgga
     3481 aagaacatgt gagcaaaaag caaagcaccg gaagaagcca acgccgcagg cgtttttcca
     3541 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa
     3601 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3661 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3721 gctttctcat agctcacgct gttggtatct cagttcggtg taggtcgttc gctccaagct
     3781 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3841 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ttggtaactg
     3901 atttagagga ctttgtcttg aagttatgca cctgttaagg ctaaactgaa agaacagatt
     3961 ttggtgagtg cggtcctcca acccacttac cttggttcaa agagttggta gctcagcgaa
     4021 ccttgagaaa accaccgttg gtagcggtgg tttttcttta tttatgagat gatgaatcaa
     4081 tcggtctatc aagtcaacga acagctattc cgttactcta gatttcagtg caatttatct
     4141 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgttagt
     4201 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4261 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4381 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     4441 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     4501 gccccagcag gcgaaaatcc tgtaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     4561 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     4621 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     4681 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     4741 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     4801 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     4861 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     4921 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     4981 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     5041 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     5101 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     5161 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     5221 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     5281 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     5341 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     5401 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata
     5461 cgactcacta ta
//
