LOCUS       sequence-14101-e        7861 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 7861)
  TITLE     Jorgensen unpublished mosSCI reagents
REFERENCE   2  (bases 1 to 7861)
  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"
COMMENT     Sequence Label: pCFJ352
FEATURES             Location/Qualifiers
     source          1..7861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1606..1630
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1776..1787)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3727..3744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3728..3744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3940..3956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3940..3956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(3963..3987)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5491..5511)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     rep_origin      complement(5648..6236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5728..5747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6407..7198,7199..7267))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7030..7049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7268..7372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7440..7458
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7496..7518)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7618..7637
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    7841..7861
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 caagtgggga tcaggaagaa gttgataaag tgattcaaaa actcgaggag ctcaacacag
       61 cgatgaaccc aattttgaga gcaacactga cgaagaacta tgacgcgagg gtaggcacta
      121 ccttgaagct atccacctgc ctgcctgccg cctatttcca tgtgcagtag gaacgtgtgt
      181 aggtataggc aggcattatc gcgtctcctt tcgaactacc tagtacctag taggtagttt
      241 gaagggaggc gcgataatgc ctgcctatgt ctacacatgt gcctactgcg catggaaata
      301 ggcggtagtc aggcaggtag atagcctcat aaaattcaat acttcctttt cagaaagaat
      361 gctggaatgt attcaaagag gagtcactct tcccattgct cagagaatat gaagagttgc
      421 tcggcgagca aaagttcatg attggagatc gatattcatg ggctgacatc gctgtcgtcg
      481 agtttctaac tcggtgtcag cagtgctacg atagcttcta tctggctcat ttcgagaggc
      541 tccgaacttt ttgccagact tttgagtctc ttccgcacat tcgtccctac attcagggac
      601 gtgttgattc atttatctaa ttttttaaaa tggatatttt tcgtttttta tttgtttgtt
      661 ttgctgataa ttcaataaag ttacgcaacg aattttagac atttatgaaa accgaaaatc
      721 gacaaaaata ttttaagaaa aatagcagca aaattcaata ttttatcaaa ttttgtttat
      781 ggtaatgaga aagaaaacat acaatatttt tctaagaaga agaaaaatga ccagaatttt
      841 aagatttttc tcgaatatac gaaaaattga taaactaaaa ttttcgattt ttccatttat
      901 tttgaaaaaa aaactcttga cggttttcac catgcgaaga ggtgagaaat attctgaaag
      961 ttatgcattt tgaaaaaatt gcaaaaaaat acgttaaaat tttttgatct attgtttttt
     1021 tttgaaaacc ggaatttaaa atggaatttt ttttcaaaaa ttaaaattaa aatttaaata
     1081 tttctagtgt ttggatagtt tttcaaaagc tagatttaac ccgcgagatg tcgaccgcta
     1141 gtgtagctta cagatggctg aagacttcgc aaaaatctta tcttagaaac ccgcgaggtg
     1201 tcggccgcta aaactttgta aacctgccaa caaaatgctt cgcattgtct gaacagatga
     1261 aaccttgaaa caaaacatat acaattgaaa tcccgtaaag tgtcgggcgc aagaactgcg
     1321 attagttgta taattggact agctcaaatt tgaccaaaat ctcattttta gacctctaag
     1381 atgtcggcca ctataacttt attcaacttg aaactgcaaa aaaacttatc aactgtatag
     1441 attcaagtat tatgtcaagt attggtttta gagctacagt acactctgat aacaaaatct
     1501 gcagaaatac ctccctgtca attcccaaaa tacctcccct catctcaatt atcccgaaaa
     1561 tcggttagtc attgccatca gaaaatcgaa ttaaaaaata aagatgcaag tttgtacaaa
     1621 aaagcaggct tgagccaatt tatccaagtc cttgtaaaaa gtattcgaaa attgttaacg
     1681 gataaaatgt ttattataat atcaaaaaca atttgtcagt tgaccacttt ttgatatatt
     1741 tgacagaaac cgggatgaat tggctcaaag ttagggcgcc cttctattac agggtttctg
     1801 ataacaaacc ggttattaac tcccaacaag ggatgatttc aattcatcat gctcaaattg
     1861 acccaaatta agttacatga caaattcatc gcccttttca actcttcttg gctcatcatc
     1921 tgttattctg ttcctattct gtctgcaccc tatacccctt tgcatactct ctcgtcattc
     1981 ctttcttgga tagtgtcttc ttctccccag ctctgctact tctatgactt gcgcccgctg
     2041 cttttccgct cgttcctctc ctctctgacg tcttcttctg cttcttttct ccttcctttt
     2101 tcccttctca atcttcctct ttttcccatt tctgtcaatc attcgaagaa gaagaagaag
     2161 aagaccctca ttcatttcat ttttttttct gtcggtgtgt gctgctcggt taagagtgag
     2221 ctctctattc cacgtcttct tctttttctt cttcgattcg aatcaatcac tccaccaaaa
     2281 cgcattcgtt tttgggattc accccgcggt tcgcaatagg tttctttttc aaattattag
     2341 cgttataaat agaaaaatgg tggagtttca ataaaaatga taatttacaa agtgtatttt
     2401 gattacatgt actcaaaaag gttgaaattt ttcataccag ttttccggaa atccatctga
     2461 tatcattatc gtatttttct ttttaaaaat gttttcaaaa aaaaacaaaa tatagctggt
     2521 tatttggcac cctctaatta ccattttcct gtcacaccac actcttttcc ttcttccact
     2581 tctttccgtt ttcagccgct tccaaccaaa ccgatatgaa agccgagcaa caacgatcga
     2641 ttccacccgg ctcggcgacc ttcccgtcgc aggtgagact cagaaaacta gagaaaccgc
     2701 tcaactaact cttgatatcc gatttcattc ttttctcttt tctttttgtt gaacttttcc
     2761 atatttccag atgccacggc caccaccaag caccgaacaa ggaatcacaa cggaatcgga
     2821 gcttgcgaag aaagctcaaa tcactccgaa cgacgtttta gcacttccgg gaatcactca
     2881 aggtatcttg ctccctttgg atttcagttt aaaacataaa tttaggattc ttatgctccc
     2941 catctgcgaa catctataac atcgagttca ccaagttcca aatccgtgat ctggacactg
     3001 agcaagtgct tttcgagatc gccaaaccgg agaacgatca ggagaatgat gagtcgccac
     3061 aggagtcggc aagatacgtg cgttatagat ttgctccaaa ctttttgaaa ctcaaaacgg
     3121 tcggagcaac tgtggaattc aaagtaggag acatcccaat ccatcatttc cgaatgatcg
     3181 aacgtcactt cttcaaagat cgccttctga agtgttttga ctttgaattc ggattctgta
     3241 ttccgaattc acgaaacaac tgtgaacata tctatgagtt ccctcaactc tctcaacaac
     3301 tcagtgagtc attattctaa aaagtacaat tcaaaagact aatctctttt tcagtggacg
     3361 acatgatcaa caatccaaac gagactcgtt ctgacagctt ctatttcgtc gataacaaac
     3421 tcgtcatgca caacaaagcc gactactcat atgatgcata aatatttaat acaaaaatgt
     3481 tctggataat tattctgtcg aatagaaaaa aaactccaaa tgtgattaaa ttccaataat
     3541 tcctgtctag tttgttcctt ccccttcccc ttctcatgtt caatgcattc ctaagctttt
     3601 cagttccccc cttgtttcta tattttttcg ctgtcctgtc acactcgcta aaaacactaa
     3661 tcacacggaa atctgttttc aataaaaact ccaactttaa ctcattttca atttcaactg
     3721 cacgggtgta aaacgacggc cagtccgtaa tacgactcac ttaaggcctt gactagaggg
     3781 taccagagct cacctaggta tctagaaccg gtgacgtcac catgggaagc ttcgtggatc
     3841 cagatatcct gcaggaattc ctcgagacgt acggtgcgcg cgatgcattc gaagatctgc
     3901 ccactagtga gtcgtattac gtagcttggc gtaatcatgg tcatagctgt ttcctgcccg
     3961 tgacccagct ttcttgtaca aagtggcagt aatcaattat ttttcgctcc cccctcctcg
     4021 caagtcggcg atttctttga agttttgtca aaagaagaga caaggacttg atagatacgc
     4081 tcatcagagt gagattcatt gcaaatagtg tgtaaatcgt ggagagacga gagaaaaggt
     4141 gtgagaagct tagaagataa agaataggaa gaagaacata tctcatttcc tattttctct
     4201 aatttttcgt tcatttttct agacattacc catttgatgg tttttcaata tggttatcag
     4261 ggtactcatt ttggtagttt tcaaagcaat ttggagtttt tttttaaatt gtcaactaca
     4321 aaaacgcagc ttaaaggtcc aaatatcata gtataaaaat tctttaaaaa attccaaatt
     4381 ttatatgatt ttttgaaaat tggaagaatg ttagtttttg ccattttttt ggaagttttt
     4441 ctccggattt tatggtactt atatttcaaa tataaatttt tgtaggaatt cgagtatgcg
     4501 agagagctat agtttactaa aaaaaagtgg caaaaactca gtaattgcca cttttttcaa
     4561 aaatgttttg aaaaatttta ctatgatatt tggccatttt ggcaccatat gtacgttact
     4621 ttttacgaaa cactctaatt tttgaaattt gagtttcatg tttaatcttt tgtataaaat
     4681 ttttatggtc cttttgtgac aggataaaaa attgcaaaaa aatagaaaac tttgtttcgt
     4741 gaaaacacct agtttaaaaa ttgtgggcca cttagcgatt ctagtcaaaa atcagacaaa
     4801 tgtcagctgc cgtaactttt tccggatcag ggttatcaaa ttttaatttt aattgtaaat
     4861 gtagaaaaat gatcggcaac attttgctaa ttgaaaatat tttgaaaatt gctatagtac
     4921 atattaatct agtattaata tagtattaat gttataatat cagttatgtg tttgaataac
     4981 tcaagcgtaa gtttccgtaa aatatcttta aattttggta attttttacg gtttggtata
     5041 aatttttgca ttttttgatt ttctacactt tttttcgatt tcgcattttc cgaatttctc
     5101 tgaaataaat cgaacaatcg gaaaattttc gagggcaccc ttagccatgc agctcaggaa
     5161 tttgacttta tgttcccatt caaatttttt taaaattaaa taaaatgcta attgtaagat
     5221 aaattacaaa aattattaaa tctcatcatc cttatgatga tagtgcagtg catgaatata
     5281 tttttccaaa taggatatga ttcgtcgagt agcttcacgt aatccgtttt cttcaatcac
     5341 catttcgaac tgcaattttt tttttggatt tttgtaacaa tattgtcata aattgtgatt
     5401 ttgatatttt gcgtagagca tcagaacctt tctgctttga attttaatgg gattaaaaat
     5461 tataaaaaac ttgcacccta attaaagcgc caactttatt atacatagtt gataattcga
     5521 tggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5581 gaaagaacat gtgagcaaaa ggccagcaaa agaccaggaa ccgtaaaaag gccgcgttgc
     5641 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5701 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5761 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5821 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5881 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5941 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6001 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6061 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6121 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6181 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6241 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6301 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6361 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6421 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6481 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6541 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6601 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6661 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6721 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6781 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6841 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6901 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6961 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7021 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7081 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7141 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7201 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7261 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7321 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7381 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     7441 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     7501 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     7561 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     7621 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     7681 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     7741 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     7801 ggcgattaag ttgggtaacg ccagggatcc gccaagctat caactttgta tagaaaagtt
     7861 g
//
