LOCUS       sequence-60689-e        7024 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7024)
  AUTHORS   Frokjaer-Jensen C, Davis MW, Sarov M, Taylor J, Flibotte S, LaBella 
            M, Pozniakovsky A, Moerman DG, Jorgensen EM
  TITLE     Random and targeted transgene insertion in Caenorhabditis elegans 
            using a modified Mos1 transposon.
  JOURNAL   Nat Methods. 2014 May;11(5):529-34. doi: 10.1038/nmeth.2889. Epub 
            2014 Mar 16.
  PUBMED    24820376
REFERENCE   2  (bases 1 to 7024)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.2889"; journalName: "Nat Methods"; date: "2014-05"; 
            volume: "11"; issue: "5"; pages: "529-34"
COMMENT     Sequence Label: pCFJ907
FEATURES             Location/Qualifiers
     source          1..7024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..160
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(201..506)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(303..322)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(826..1506)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     1421..1440
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1560..1566,1567..1584,1585..1590))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1615..1738)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1746..1763)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1746..1762)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2579..3373
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2633..2652)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3243..3262
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4074..4090
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4075..4091
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4746..4768
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4806..4824)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4892..4996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4997..5065,5066..5857)
                     /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     complement(5215..5234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6028..6616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6517..6536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaac
       61 gagaaacgta aaatgatata aatatcaata tattaaatta gattttgcat aaaaaacaga
      121 ctacataata ctgtaaaaca caacatatcc agtcactatg gtcgacctgc agactggctg
      181 tgtataaggg agcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
      241 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
      301 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
      361 tcacggggga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
      421 ccgggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
      481 ttataggtgt aaaccttaaa ctgcatttca ccagcccctg ttctcgtcgg caaaagagcc
      541 gttcatttca ataaaccggg cgacctcagc catcccttcc tgattttccg ctttccagcg
      601 ttcggcacgc agacgacggg cttcattctg catggttgtg cttaccgaac cggagatatt
      661 gacatcatat atgccttgag caactgatag ctgtcgctgt caactgtcac tgtaatacgc
      721 tgcttcatag catacctctt tttgacatac ttcgggtata catatcagta tatattctta
      781 taccgcaaaa atcagcgcgc aaatacgcat actgttatct ggcttttagt aagccggatc
      841 ctctagatta cgccccgcct gccactcatc gcagtactgt tgtaattcat taagcattct
      901 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
      961 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     1021 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     1081 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1141 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     1201 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     1261 cagctcaccg tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag
     1321 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     1381 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     1441 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     1501 ctccatttta gcttccttag ctcctgaaaa tctcgacgga tcctaactca aaatccacac
     1561 attatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt
     1621 gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     1681 aatatattga tatttatatc attttacgtt tctcgttcaa ctttattata catagttgat
     1741 aattcactgg ccgtcgtttt acaggccgga tgggccatgg caccatttct aaattttagt
     1801 ttaataaaaa aattttaaaa taatacgagt tttgagacta actttcaata aaaaccaaaa
     1861 aattgcttca aaaaagagct cgcgccgatt tacgataaac acagcgcgct acgacactcc
     1921 gaaaaccgcg ccgcaccatt acattctccg ccccaccgca cggtttgcgg aaggcgatga
     1981 aattgctggg cctaaaattt taaccctttt tttcccgttt ttcaactgtt ttcccgattt
     2041 tttcgatatt tttttcaggt ttttagttag aaaagtatat aaaaattgca ttatttgtag
     2101 ttttcgctcg taatttgtga tagaaaacgg ccaatttgcc tagatttgtg attctaccaa
     2161 gtggaatttt tgggaaaata tcaatattcc acgattttta ttcaaatcat aacgtttcga
     2221 aattatcagc ttaaataaaa ctaaaataaa taataaaata ctaaaaaccc tgaatttctc
     2281 ggttctcggt tcagcttgag ttcgttctat acacgttatc gatttttgtc atttcgttta
     2341 atcatgggtt ttgattcatt ttaaatatga aatttgattt attcctcgtt tcgaagttgg
     2401 tttgtcttgt ccggccggta aacaaatgtt attaaaaaaa ataataaatt tcgaaatttt
     2461 tcgtaatgtt ccgcaatcgt gtttttccgt ttatctcgtc ttcccttcct tcaaatatac
     2521 tatttatttt ctcaattcac atattttcag ccagccctgc tcaacaagtg gaataaaaat
     2581 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2641 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2701 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2761 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2821 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2881 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2941 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3001 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3061 gcatcagggg ctcgcaccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3121 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3181 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3241 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3301 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3361 cgagttcttc tgagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     3421 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     3481 ttttaagcca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     3541 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     3601 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     3661 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     3721 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     3781 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     3841 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     3901 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     3961 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     4021 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttctcgaggt
     4081 cgacggtatc gataagcttg atccggatct gcggcttact caccagacct ggccccatcc
     4141 gattaccacc tattcgcttc gatgggacac gcactcgctg agcagcgctt cgattcttac
     4201 gaaagtgtga aaaaatggct cgatgaatgg ttcgccgcaa aagacgatga gttctactgg
     4261 cgtggaatcc acaaattgcc cgagagatgg gaaaaatgtg tagctagcga cggcaaatac
     4321 tttgaataaa tgattttttc tttttccaca aaatttaacg tgttttttga tttaaaaaaa
     4381 acgacatttc atacttgtac acctgatatc tagataccta ggtgagctct ggtacccccc
     4441 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4501 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
     4561 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     4621 tctcagtgca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4681 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4741 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4801 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4861 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4921 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4981 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5041 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5101 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5161 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5221 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5281 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5341 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5401 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5461 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5521 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5581 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5641 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5701 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5761 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5821 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5881 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5941 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     6181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg tcttttgctg
     6661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6721 cgcctttgag tgagctgata ccgcaccgta cgtctcgagg aattctatca ggtgtacaag
     6781 tatgaaatgt cgtttcgaac atatagatgt ctcgcaaacg taaatattta tcgattgtca
     6841 taaaactttg accttgtgaa gtgtcaacct tgactgtcga accaccatag tttggcgcga
     6901 attgagcgtc ataattgttt actctcagtg cagtcaacat gtcgagtttc gtgccgaata
     6961 aagagcaaac gcggacagta ttaattttct gttttcattt gaagaaaaca gctgcggccg
     7021 gatc
//
