LOCUS       sequence-137186-        6017 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 6017)
  AUTHORS   King BC, Nowakowska J, Karsten CM, Kohl J, Renstrom E, Blom AM
  TITLE     Truncated and full-length thioredoxin-1 have opposing activating and
            inhibitory properties for human complement with relevance to 
            endothelial surfaces.
  JOURNAL   J Immunol. 2012 Apr 15;188(8):4103-12. Epub 2012 Mar 19.
  PUBMED    22430737
REFERENCE   2  (bases 1 to 6017)
  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 
            Immunol."; date: "2012-04-15"; volume: "188(8)"; pages: "4103-12. 
            Epub 2012 Mar 19"
COMMENT     Sequence Label: pET16b-Trx
FEATURES             Location/Qualifiers
     source          1..6017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        40..58
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    59..83
                     /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             98..120
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             133..162
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             175..186
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(558..576)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      572..619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(794..799,800..816,817..822))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(849..867)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        935..1039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1040..1108,1109..1900)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1258..1277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2071..2659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2560..2579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2813..2830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2845..2984
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2910..2929)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3070..3092
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4588..5670)
                     /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     5632..5651
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5671..5748)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5954..5973
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
       61 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
      121 tataccatgg gccatcatca tcatcatcat catcatcatc acagcagcgg ccatatcgaa
      181 ggtcgtcata tggtgaagca gatcgagagc aagactgctt ttcaggaagc cttggacgct
      241 gcaggtgata aacttgtagt agttgacttc tcagccacgt ggtgtgggcc ttgcaaaatg
      301 atcaagcctt tctttcattc cctctctgaa aagtattcca acgtgatatt ccttgaagta
      361 gatgtggatg actgtcagga tgttgcttca gagtgtgaag tcaaatgcat gccaacattc
      421 cagtttttta agaagggaca aaaggtgggt gaattttctg gagccaataa ggaaaagctt
      481 gaagccacca ttaatgaatt agtctaagga tccggctgct aacaaagccc gaaaggaagc
      541 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      601 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg caagaggccc
      661 ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg
      721 acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt
      781 gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga attcttgaag
      841 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
      901 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1261 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     1321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1561 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1921 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1981 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2041 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2101 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2161 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     2221 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2281 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2341 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2401 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2461 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2521 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2581 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2641 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2701 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2761 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2821 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2881 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2941 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3001 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3061 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3121 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3181 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3241 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     3301 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     3361 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     3421 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     3481 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     3541 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     3601 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     3661 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     3721 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     3781 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     3841 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     3901 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     3961 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     4021 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
     4081 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     4141 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     4201 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     4261 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     4321 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     4381 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     4441 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     4501 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     4561 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4621 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4681 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     4741 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     4801 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     4861 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     4921 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     4981 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     5041 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     5101 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     5161 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     5221 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     5281 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     5341 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     5401 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     5461 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     5521 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     5581 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     5641 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     5701 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     5761 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     5821 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5881 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5941 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6001 tgatgccggc cacgatg
//
