LOCUS       sequence-12672-e        6282 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 6282)
  AUTHORS   Smith SP, Barber KR, Dunn SD, Shaw GS
  TITLE     Structural influence of cation binding to recombinant human brain 
            S100b: evidence for calcium-induced exposure of a hydrophobic 
            surface.
  JOURNAL   Biochemistry. 1996 Jul 9. 35(27):8805-14.
  PUBMED    8688416
REFERENCE   2  (bases 1 to 6282)
  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: 
            "Biochemistry. 1996 Jul 9. 35(27):8805-14."
COMMENT     Sequence Label: pSS2 (hS100B)
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..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)."
     primer_bind     16..38
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     regulatory      83..92
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(704..721)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      874..960
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1052..1079
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1098..1189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1190..1258,1259..2050)
                     /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(1408..1427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2221..2809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2710..2729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2963..2980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2995..3135
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3061..3080)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3221..3243
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             3287..4369
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3306..3325)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     5990..6009
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6257..6262,6263..6278,6279..6282,1..3)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagaattct ggaggatttt
       61 aaaatgtctg agctggagaa ggccatggtg gccctcatcg acgttttcca ccaatattct
      121 ggaagggagg gagacaagca caagctgaag aaatccgaac tcaaggagct catcaacaat
      181 gagctttccc atttcttaga ggaaatcaaa gagcaggagg ttgtggacaa agtcatggaa
      241 acactggaca atgatggaga cggcgaatgt gacttccagg aattcatggc ctttgttgcc
      301 atggttacta ctgcctgcca cgagttcttt gaacatgagt gagattagaa agcagccaaa
      361 cctttcctgt aacagagacg gtcatgcaag aaagcagaca gcaagggctt gcagcctagt
      421 aggagctgag ctttccagcc gtgttgtagc taattaggaa gcttggacca aaaaagcgat
      481 gtaacaccgg cttgaaaagc acaaaagcca gtctggaaac aggctggctt ttttttgcgc
      541 gtgtgacccg tcctgaatag cgttcacata gatcctgctg atataaaacc cccctgtttt
      601 cctgtttatt cattgatcga aataagagca aaaacatcca cctgacgctt aaattaaggt
      661 accaagctag cttctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
      721 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
      781 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
      841 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
      901 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      961 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     1021 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
     1081 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc
     1441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2341 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     2401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2581 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg
     2641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2881 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2941 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
     3001 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     3061 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3121 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3181 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3241 aggttttcac cgtcatcacc gaaacgcgcg aggcagggtg gtgaatgtga aaccagtaac
     3301 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     3361 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     3421 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     3481 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     3541 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     3601 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     3661 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     3721 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     3781 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     3841 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     3901 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     3961 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     4021 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     4081 tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     4141 gtcaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     4201 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     4261 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4321 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcac
     4381 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     4441 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4501 tgttaagggc ggttttttcc tgtttggtca cttgatgcct ccgtgtaagg gggaatttct
     4561 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     4621 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     4681 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     4741 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     4801 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     4861 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     4921 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     4981 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct
     5041 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     5101 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     5161 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     5221 cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag
     5281 gcggcataaa tcgccgtgac gatcagcggt ccagtgatcg aagttaggct ggtaagagcc
     5341 gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc
     5401 tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc
     5461 cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg
     5521 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     5581 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     5641 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5701 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5761 ctgactgggt tgaaggctct caagggcatc ggtcgacgct ctcccttatg cgactcctgc
     5821 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     5881 gcatgcaagg agatggcgcc caacagtccc ccggccacgg gcctgccacc atacccacgc
     5941 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6001 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6061 cgtagaggat ccggagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     6121 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     6181 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     6241 attctgaaat gagctgttga caattaatca tcggctcgta ta
//
