LOCUS       sequence-8982-ex        9549 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 9549)
  AUTHORS   Christian Frøkjær-Jensen, M Wayne Davis, Christopher E Hopkins, 
  TITLE     Single-copy insertion of transgenes in Caenorhabditis elegans
  PUBMED    18953339
REFERENCE   2  (bases 1 to 9549)
  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: pCFJ68 - Exp5605_Punc-122::GFP::unc-54utr
FEATURES             Location/Qualifiers
     source          1..9549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1437..1461
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1783..1811)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2559..2579
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             5588..5599
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5750..5774)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7113..7133)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(7153..7169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7153..7169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7166..7188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(7315..7903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7395..7414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8074..8865,8866..8934))
                     /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     8697..8716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8935..9039)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9107..9125
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     9285..9304
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9498..9520
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9512..9529
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9513..9529
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(9537..9549,1..8)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
ORIGIN
        1 taaagttgct gtttttgaac acggcgatat gtatctgtag atcatcccca attttcaaca
       61 atgaatctaa atactgaact ttttgtgtaa tctttagtgc agaaaaatga ccaaaatcag
      121 cgattttccg attataccca aaatatctca gctcaatgag cacctataaa aatgtaaaaa
      181 taaagaaagt agtcgaacca tattttgtaa aggctaaaaa actgaataaa tttttatcca
      241 tccagcgcgg cgttacaatc cgtggcccat gtcacgcacg gctttcaaag aaaaaaaaac
      301 agtctgtttg cgcgacatcg atctggcaaa aatcctaaaa acgactatgc gtggctcact
      361 ttatattaat atgaaaaata caaacttggt ggtgcaggtg ctgcagaggc catcgatgaa
      421 gaagtggagg ccttgttgtc gatctacaaa attgtatggt tgtttgagat tttccttgaa
      481 acgctctcaa tcttggatat gatataaatt ctggataaga tatgatggat atgatctatg
      541 tttgatagag tgcatttgaa aggataacgg ccctgcagac atttttgaaa aaataacaaa
      601 catttaattt tctcactcgt ttaggctatt cccctatttt tgatattcct tcgcacatat
      661 gaaaactact ttttttcgaa actgttaact ccagaattta taaatctata gcccttactt
      721 gattatttat tatcatggtt actcacctca tgtctctttc tcttttcaag aagtcttcgt
      781 tcatatctgt tgtctctttc ctccttttga atctctgctt tgctcttcgc cattgtttcc
      841 tgaaaataat gtaacttgaa ttgtgtaata tactttttta atttgaattt ggcttgtaac
      901 gcggaatcac tacgtgcggg atcatttctt actagaaaac ccagaaaatg ccatatttca
      961 cttatctcgg ggtcatttct aattagaaaa gctacaaaaa ccatttctaa ttacggtgac
     1021 tttagaactg gtcgaaatag ctaattagaa atgggttttt gctaattaga ggtgactcta
     1081 tgttatctga gtaagcctcg tttgtatgta aactcacagt actaataaat gcaagacacc
     1141 cgggtttgtc tagatatgaa ataattgaaa tatcaattct gacagacaat aatggtaatc
     1201 ttgataagga gttccacgcc caggagaaca cgttagtttt cttgtttttg attgcgtgcg
     1261 ttattttgga gaaaaactcg attttttaca aaataatttt ttgaaaggaa cactgttcaa
     1321 taagttttgt cttttttctc agttgtgata cggtttttta ttcttttttg tagttataca
     1381 gaagaccgtt acgaaacaga ctgatatcga aacaaacact gatcttactt gcacttacca
     1441 ctttgtacaa gaaagctggg tgacacgcag tttccctcat ttctcatcct tcacgcaata
     1501 tttggagact tattctaaac atttccaaaa ccccacggaa atgactatca gtcaacaatt
     1561 tgtttattct acgtgcatat catattacca ctatattctg gcaactagca tgaaatttta
     1621 cttcattggg ctcacaatat ggttgcgaca ggatccccgg gattggccaa aggacccaaa
     1681 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
     1741 ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
     1801 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
     1861 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
     1921 tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca
     1981 cttgtcacta ctttctgtta tggtgttcaa tgcttctcga gatacccaga tcatatgaaa
     2041 cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
     2101 ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca
     2161 tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     2221 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     2281 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     2341 gttgtaagtt taaacttgga cttactaact aacggattat atttaaattt tcagaacttc
     2401 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat
     2461 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     2521 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     2581 gctgctggga ttacacatgg catggatgaa ctatacaaat agcattcgta gaattccaac
     2641 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     2701 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     2761 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     2821 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     2881 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     2941 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     3001 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     3061 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     3121 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     3181 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     3241 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     3301 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     3361 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     3421 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     3481 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3541 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3601 agatctgccc gggcggccgc ctagttctag acattctcta atgaaaaaat ctttcagttg
     3661 aaattgaaaa tgagttaaag ttggagtttt tattgaaaac agatttccgt gtgattagtg
     3721 tttttagcga gtgtgacagg acagcgaaaa aatatagaaa caagggggga actgaaaagc
     3781 ttaggaatgc attgaacatg agaaggggaa ggggaaggaa caaactagac aggaattatt
     3841 ggaatttaat cacatttgga gttttttttc tattcgacag aataattatc cagaacattt
     3901 ttgtattaaa tatttatgca tcatatgagt agtcggcttt gttgtgcatg acgagtttgt
     3961 tatcgacgaa atagaagctg tcagaacgag tctcgtttgg attgttgatc atgtcgtcca
     4021 ctgaaaaaga gattagtctt ttgaattgta ctttttagaa taatgactca ctgagttgtt
     4081 gagagagttg agggaactca tagatatgtt cacagttgtt tcgtgaattc ggaatacaga
     4141 atccgaattc aaagtcaaaa cacttcagaa ggcgatcttt gaagaagtga cgttcgatca
     4201 ttcggaaatg atggattggg atgtctccta ctttgaattc cacagttgct ccgaccgttt
     4261 tgagtttcaa aaagtttgga gcaaatctat aacgcacgta tcttgccgac tcctgtggcg
     4321 actcatcatt ctcctgatcg ttctccggtt tggcgatctc gaaaagcact tgctcagtgt
     4381 ccagatcacg gatttggaac ttggtgaact cgatgttata gatgttcgca gatggggagc
     4441 ataagaatcc taaatttatg ttttaaactg aaatccaaag ggagcaagat accttgagtg
     4501 attcccggaa gtgctaaaac gtcgttcgga gtgatttgag ctttcttcgc aagctccgat
     4561 tccgttgtga ttccttgttc ggtgcttggt ggtggccgtg gcatctggaa atatggaaaa
     4621 gttcaacaaa aagaaaagag aaaagaatga aatcggatat caagagttag ttgagcggtt
     4681 tctctagttt tctgagtctc acctgcgacg ggaaggtcgc cgagccgggt ggaatcgatc
     4741 gttgttgctc ggctttcata tcggtttggt tggaagcggc tgaaaacgga aagaagtgga
     4801 agaaggaaaa gagtgtggtg tgacaggaaa atggtaatta gagggtgcca aataaccagc
     4861 tatattttgt ttttttttga aaacattttt aaaaagaaaa atacgataat gatatcagat
     4921 ggatttccgg aaaactggta tgaaaaattt caaccttttt gagtacatgt aatcaaaata
     4981 cactttgtaa attatcattt ttattgaaac tccaccattt ttctatttat aacgctaata
     5041 atttgaaaaa gaaacctgtt gcgaaccgcg gggtgaatcc caaaaacgaa tgcgttttgg
     5101 tggagtgatt gattcgaatc gaagaagaaa aagaagaaga cgtggaatag agagctcact
     5161 cttaaccgag cagcacacac cgacagaaaa aaaaatgaaa tgaatgaggg tcttcttctt
     5221 cttcttcttc gaatgattga cagaaatggg aaaaagagga agattgagaa gggaaaaagg
     5281 aaggagaaaa gaagcagaag aagacgtcag agaggagagg aacgagcgga aaagcagcgg
     5341 gcgcaagtca tagaagtagc agagctgggg agaagaagac actatccaag aaaggaatga
     5401 cgagagagta tgcaaagggg tatagggtgc agacagaata ggaacagaat aacagatgat
     5461 gagccaagaa gagttgaaaa gggcgatgaa tttgtcatgt aacttaattt gggtcaattt
     5521 gagcatgatg aattgaaatc atcccttgtt gggagttaat aaccggtttg ttatcagaaa
     5581 ccctgtaata gaagggcgcc ctaactttga gccaattcat cccggtttct gtcaaatata
     5641 tcaaaaagtg gtcaactgac aaattgtttt tgatattata ataaacattt tatccgttaa
     5701 caattttcga atacttttta caaggacttg gataaattgg ctcaaagtaa gcctgctttt
     5761 ttgtacaaac ttgcacgaag tgagtttgct accatcatgg aatcaggagg acatttcgac
     5821 aatgtcatat aatgtttgaa aaatgatttt aacaagggtc tcattcgaga taagcaatgc
     5881 tctgagcaag tggctcgtcg tcttcaagaa tctcttgttg atattcattt ggatcctgtt
     5941 ttgcacgaag catgtgccat ggatattcag agtttttgca aaaaagttcc gccaggtcaa
     6001 acaatctttt taatttttat atgatacttc gggtgtattt ttcttctttt tcctgttcat
     6061 tttcgctttt ttctcacttt tttgtaacaa aatctctcac ttttctgtaa gtcttaacaa
     6121 aagttcaatt ggctcgatca agtaaaattt cggccgtatg aacgttttca aaggttttca
     6181 ttttttttta agttcattca caatatcatg aatctctcgt cctttggttt ccggtaagta
     6241 gcgaaataat ataagcatac ttataaatga aggaattgaa tatagaataa ggaaagcata
     6301 actaccaata acggaatata gaggtaatac tgtaaatgta ctgataacca ccattacagt
     6361 attcattgaa tatgccacag attgagcaat actacgatgt ttttgaggta ccagttcaga
     6421 ggatataaac cacgaaattg gaccaactcc agttctgaaa tttcattatt cgattaaatc
     6481 aaaatatatt tgaaaaacct cacccataag tataaccgta gatgatcaat gcaactaggc
     6541 atccgtattt catcgggtca atcagaatac gtatttttgc gaagaatacg aatgcaatta
     6601 gagctaaatt attgagaaat gtgaagagaa ggagaagctt gcgtcgacca actctgtaaa
     6661 cttattcata caattttgtc ttcaaatttt gaacgtttaa aactatctta ttcaatgagt
     6721 tagcgggttt ttatagtgag ttagttcacg attatctaat aaaactatgt ccaaacatcc
     6781 taactgcgac cgtattctca ctaactcgtc ttaaaaacgt tgagattatt ttaaaccctt
     6841 cttcagtaca actacagtac tgatgcacaa atacaataat aataatacat tcagacatac
     6901 tctgaacaat gtaacttgtt tttgttatac tgtggtaaca tttattgaaa tgaaatataa
     6961 ccttactttt caatcacaat tccaccagta attgttccag aaacataagc taaacccatt
     7021 gcagttgtac tccattcagc aatttctgac tcaacatttg cattttctag aaagaaagta
     7081 gaggataaaa gaagagccca taatccgaca agcaactttt ctatacaaag ttgatagctt
     7141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctgg tatcagctca
     7201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7261 agcaaaaggc cagcaaaaga ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7741 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9061 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9121 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9181 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9241 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9301 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9361 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9421 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9481 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattatcaac
     9541 tatgtataa
//
