LOCUS       sequence-37066-e        7635 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 7635)
  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 7635)
  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: CD4d3+4-blac
FEATURES             Location/Qualifiers
     source          1..7635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             907..1695
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     polyA_signal    1730..1785
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1784..1803)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4021..4039)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4107..4211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4212..4280,4281..5072)
                     /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"
     rep_origin      5243..5831
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5732..5751
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5985..6002
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6119..6140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6155..6160,6161..6178,6179..6185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6193..6209
                     /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     6198..6220
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6217..6233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6217..6233
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        6504..6883
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6884..7087
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7038..7058
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7084..7108
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     7589..7608
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gcggccgctc cggctagcga taagcttgac tcagatcccc atccgctcaa gcaggccacc
       61 atgtgccgag gcttctcttt caggcacttg ctgccgctgc tgctcctgca gctgtcaaaa
      121 ctcctagttg tcacccaagg gtcgacatcc atcacggcct ataagagtga gggggagtca
      181 gcggagttct ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg
      241 aaggcagaga aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag
      301 gtgtctgtgc agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc
      361 acccttcaga taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact
      421 ctggacagag ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc
      481 gacagcaaca ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc
      541 ttgaagcagg agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag
      601 gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac
      661 tccaagatcc aggttttatc caaagggttg aattccggcg gcggctccgg aggcggtacc
      721 ggtggcggcg acctagcccc acagatgctt cgagaactcc aggagactaa tgcggcgctg
      781 caagacgtga gagagctctt gcgacagcag gtcaaggaga tcaccttcct gaagaatacg
      841 gtgatggaat gtgacgcctg cggtctagac agaaacctgc ctccactagc acctctgggc
      901 ccacacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      961 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1021 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1081 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1141 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1201 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1261 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1321 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1381 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1441 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1501 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1561 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1621 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1681 attaagcatt ggtaattaat taaggatccc ccgacctcga cctctggcta ataaaggaaa
     1741 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     1801 gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg atcgatcccc
     1861 gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat
     1921 gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac atgtgacacg
     1981 gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat aaatggatgt
     2041 gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt ctgtggactg
     2101 caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca atgctggggg
     2161 acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt caatcagagg
     2221 ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag tgtttataag
     2281 gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta tatactatcc
     2341 agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct atcgaattag
     2401 ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct gtcacggttt
     2461 tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac aagggcagtg
     2521 gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt cttcattctc
     2581 cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat gtgaggtgct
     2641 cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg ttcagtggtg
     2701 gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa atactagtgt
     2761 aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg gacaagccgt
     2821 aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata atcctagtgc
     2881 aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag gtgaaccatg
     2941 ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca gcggactcca
     3001 ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg ggcccataaa
     3061 caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc gtcagccccc
     3121 acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg ctgattgtaa
     3181 ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc accccgggga
     3241 atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg atctggagga
     3301 caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct catattcacg
     3361 aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta cccaaatatc
     3421 tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     3481 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     3541 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     3601 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct aatagagatt
     3661 agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca aatatctgga
     3721 tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     3781 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     3841 tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg
     3901 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg
     3961 tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga agacgaaagg
     4021 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4081 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4141 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4201 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4261 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4321 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4381 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4441 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4501 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4561 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4621 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4681 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4741 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4801 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4861 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4921 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4981 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5041 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5101 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5161 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5881 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5941 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6001 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6061 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6121 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     6181 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     6241 acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat catcgcagat
     6301 ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga agatctatac
     6361 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     6421 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     6481 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
     6541 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6601 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6661 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6721 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     6781 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     6841 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     6901 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     6961 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     7021 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     7081 cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg tctgcgaggg
     7141 ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt actcttggat
     7201 cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc gagtccgcat
     7261 cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg caaggtaggc
     7321 tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg gaggtgctgc
     7381 tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg aggtgtggca
     7441 ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat tacttctgcg
     7501 ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc cgatctggcc
     7561 atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt ccactcccag
     7621 gtccaagttt aaact
//
