LOCUS       sequence-37198-e        7131 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 7131)
  AUTHORS   Bushell KM, Sollner C, Schuster-Boeckler B, Bateman A, Wright GJ
  TITLE     Large-scale screening for novel low-affinity extracellular protein 
            interactions.
  JOURNAL   Genome Res. 2008 Apr;18(4):622-30. Epub 2008 Feb 22.
  PUBMED    18296487
REFERENCE   2  (bases 1 to 7131)
  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: "Genome 
            Res."; date: "2008-04"; volume: "18(4)"; pages: "622-30. Epub 2008 
            Feb 22"
COMMENT     Sequence Label: secretedBirA-8his
FEATURES             Location/Qualifiers
     source          1..7131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1239..1258
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1447..2409
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1447..2409
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2416..2439
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    2511..2566
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2565..2584)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4802..4820)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4888..4992
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4993..5061,5062..5853)
                     /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(5211..5230)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6024..6612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6513..6532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6766..6783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6900..6921
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6936..6941,6942..6959,6960..6966)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6974..6990
                     /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     6979..7001
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6998..7014
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6998..7014
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
       61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
      781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
      841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
      901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
      961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     1261 ccactcccag gtccaagttt aaacggatct ctagcgaatt ccctctagag ggcccgtttc
     1321 tgctagcctc gacactcaga tccccatccg ctcaagcagg ccaccatgtg ccgaggcttc
     1381 tctttcaggc acttgctgcc gctgctgctc ctgcagctgt caaaactcct agttgtcacc
     1441 caagggatga aggataacac cgtgccactg aaattgattg ccctgttagc gaacggtgaa
     1501 tttcactctg gcgagcagtt gggtgaaacg ctgggaatga gccgggcggc tattaataaa
     1561 cacattcaga cactgcgtga ctggggcgtt gatgtcttta ccgttccggg taaaggatac
     1621 agcctgcctg agcctatcca gttacttaat gctaaacaga tattgggtca gctggatggc
     1681 ggtagtgtag ccgtgctgcc agtgattgac tccacgaatc agtaccttct tgatcgtatc
     1741 ggagagctta aatcgggcga tgcttgcatt gcagaatacc agcaggctgg tcgtggtcgc
     1801 cggggtcgga aatggttttc gccttttggc gcaaacttat atttgtcgat gttctggcgt
     1861 ctggaacaag gcccggcggc ggcgattggt ttaagtctgg ttatcggtat cgtgatggcg
     1921 gaagtattac gcaagctggg tgcagataaa gttcgtgtta aatggcctaa tgacctctat
     1981 ctgcaggatc gcaagctggc aggcattctg gtggagctga ctggcaaaac tggtgatgcg
     2041 gcgcaaatag tcattggagc cgggatcaac atggcaatgc gccgtgttga agagagtgtc
     2101 gttaatcagg ggtggatcac gctgcaggaa gcggggatca atctcgatcg taatacgttg
     2161 gcggccatgc taatacgtga attacgtgct gcgttggaac tcttcgaaca agaaggattg
     2221 gcaccttatc tgtcgcgctg ggaaaagctg gataatttta ttaatcgccc agtgaaactt
     2281 atcattggtg ataaagaaat atttggcatt tcacgcggaa tagacaaaca gggggcttta
     2341 ttacttgagc aggatggaat aataaaaccc tggatgggcg gtgaaatatc cctgcgtagt
     2401 gcagaaaaac tcgagcacca ccaccaccac caccaccact gagtctagag tcgacccggg
     2461 cggccgctcg aggccggcaa ggccggatcc cccgacctcg acctctggct aataaaggaa
     2521 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2581 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2641 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2701 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2761 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2821 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2881 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2941 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3001 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3061 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3121 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3181 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3241 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3301 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3361 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3421 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3481 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3541 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3601 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3661 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3721 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3781 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3841 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3901 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3961 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4021 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4081 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4141 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4201 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4261 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4321 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4381 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4441 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4501 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4561 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4621 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4681 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4741 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4801 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4861 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4921 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4981 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5041 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5101 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5161 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5221 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5281 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5341 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5401 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5461 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5521 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5581 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5641 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5701 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5761 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5821 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5881 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5941 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6061 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6781 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6841 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6901 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6961 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7021 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     7081 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg a
//
