LOCUS       sequence-271191-        6161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  salL gene from S. tropica CNB-440 ligated into pHIS8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6161)
  AUTHORS   Eustaquio AS, Pojer F, Noel JP, Moore BS
  TITLE     Discovery and characterization of a marine bacterial SAM-dependent 
            chlorinase.
  JOURNAL   Nat Chem Biol. 2008 Jan;4(1):69-74. doi: 10.1038/nchembio.2007.56. 
            Epub 2007 Dec 2.
  PUBMED    18059261
REFERENCE   2  (bases 1 to 6161)
  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: 
            "10.1038/nchembio.2007.56"; journalName: "Nat Chem Biol"; date: 
            "2008-01"; volume: "4"; issue: "1"; pages: "69-74"
COMMENT     Sequence Label: pAEM7
FEATURES             Location/Qualifiers
     source          1..6161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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             79..101
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             114..137
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             144..161
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1046..1063
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1116..1134)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1130..1177
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1214..1669
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1301..1320)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1511..1532
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1761..2576)
                     /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(1761..2576)
                     /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     2484..2503
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2568..2587
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2698..3286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3187..3206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3440..3457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3472..3611
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3537..3556)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3697..3719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3713..3904)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4713..5795)
                     /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     5757..5776
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5796..5873)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6079..6098
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg aaacaccacc
      121 accaccacca ccaccacggt ggtctggttc cgcgtggttc ccatggcatg cagcacaatc
      181 tcattgcctt cctctccgac gtcgggtccg ccgacgaagc ccacgcactc tgcaagggag
      241 tcatgtacgg cgtcgcgccg gcagcgacga tcgtggacat aacccatgac gtcgcgccct
      301 ttgatgtccg ggaaggcgca ctgtttctcg ctgatgtccc gcactccttc ccggctcata
      361 cggtcatctg cgcatacgtc tatcccgaga ccggcacggc gacccacacc attgcagtgc
      421 gcaatgagaa gggtcaactg ctcgtcggac ccaacaatgg gctgctgagc ttcgcactcg
      481 acgcctcccc tgcggtcgag tgtcacgagg tcctctcccc cgatgtcatg aaccagccgg
      541 tgacgccaac ctggtacggc aaggacatcg tcgctgcctg cgccgcccac ctcgctgccg
      601 gcaccgacct tgccgccgtc ggcccccgga tcgacccgaa gcagatcgtt cggctgccct
      661 atgcctccgc cagcgaggtg gagggcggga tccgcgggga ggtggtacgc atcgaccggg
      721 ctttcggcaa cgtatggacc aacataccca cccacttgat tggctcgatg ctgcaggacg
      781 gggagcgact ggaagtcaag atcgaagccc tcagcgacac ggtgctcgag ctgcccttct
      841 gcaaaacctt cggcgaggtc gacgaggggc agccactgct gtacctcaac agtcggggcc
      901 ggcttgccct cgggctcaac cagtccaact tcatcgagaa gtggcctgtc gtacccggtg
      961 acagcatcac cgtctcccca agggtccccg atagtaacct gggcccggtg ctcgggtagc
     1021 tgacaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa
     1081 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1141 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1201 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1261 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1321 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1381 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1441 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1501 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1561 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1621 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     1681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1741 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     1801 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     1861 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1921 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1981 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     2041 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     2101 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     2161 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     2221 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     2281 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     2341 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     2401 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     2461 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     2521 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
     2581 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
     2641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2821 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     3481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     3541 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     3601 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3721 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3781 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3841 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3901 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     3961 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     4021 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     4081 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     4141 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     4201 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     4261 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     4321 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca
     4381 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     4441 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     4501 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     4561 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     4621 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     4681 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4741 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4801 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     4861 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     4921 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     4981 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5041 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5101 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5161 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     5221 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     5281 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     5341 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     5401 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     5461 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     5521 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     5581 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     5641 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     5701 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     5761 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     5821 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     5881 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     5941 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     6001 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6061 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6121 gccggtgatg ccggccacga tgcgtccggc gtagaggatc g
//
