LOCUS       sequence-215667-        4436 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 4436)
  AUTHORS   Krajcikova D, Hartley RW, Sevcik J
  TITLE     Isolation and purification of two novel streptomycete RNase 
            inhibitors, SaI14 and SaI20, and cloning, sequencing, and expression
            in Escherichia coli of the gene coding for SaI14.
  JOURNAL   J Bacteriol 1998 Mar;180(6):1582-5.
  PUBMED    9515932
REFERENCE   2  (bases 1 to 4436)
  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: "J 
            Bacteriol"; date: "1998-03"; volume: "180(6)"; pages: "1582-5"
COMMENT     Sequence Label: pSaI20
FEATURES             Location/Qualifiers
     source          1..4436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    4..20
                     /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     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(394..411)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      564..650
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      742..769
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        788..879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(880..948,949..1740)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1098..1117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1911..2499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2400..2419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2685..2825
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2751..2770)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2911..2933
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3011..3088
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3089..4171
                     /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(3108..3127)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4403..4408,4409..4425,4426..4432)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 gaattgtgag cggataacaa tttcacacag gaaacagacc atggcggaca acgagctgat
       61 cgtcgacctt cggggccggc agatcgagac actgaacgac ttctgggacg cagtgagcga
      121 gccctgcggg ttgccggagt ggttcggccg gaatttggac gcgtggtcgg acacgatcga
      181 gacccggggc atctccgagg tgatcgacag ccacgacatc ctggtcgtgc acgtcgacca
      241 gcggggtctc ttcgaagggc accggaggga agccgacgtc ctggctgaca ccttcgacgg
      301 agagcagaac cagctcatcg tccacggacc ggattggcct gcggcaacgg attgaaagct
      361 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
      421 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
      481 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
      541 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      601 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      661 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      721 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
      781 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1081 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2041 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2701 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     2761 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2821 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2881 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2941 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     3001 catttacgtt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     3061 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     3121 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     3181 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     3241 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     3301 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     3361 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     3421 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     3481 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     3541 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     3601 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     3661 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     3721 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     3781 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     3841 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     3901 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     3961 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     4021 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     4081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg
     4201 aattgatctg gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     4261 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     4321 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     4381 gcaaatattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtg
//
