LOCUS       sequence-174001-        6359 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 6359)
  AUTHORS   Merritt C, Rasoloson D, Ko D, Seydoux G
  TITLE     3' UTRs are the primary regulators of gene expression in the C. 
            elegans germline.
  JOURNAL   Curr Biol. 2008 Oct 14;18(19):1476-82. doi: 
            10.1016/j.cub.2008.08.013. Epub 2008 Sep 25.
  PUBMED    18818082
REFERENCE   2  (bases 1 to 6359)
  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"; doi: 
            "10.1016/j.cub.2008.08.013"; journalName: "Curr Biol"; date: 
            "2008-10-14- 14"; volume: "18"; issue: "19"; pages: "1476-82"
COMMENT     Sequence Label: pCG150
FEATURES             Location/Qualifiers
     source          1..6359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..184
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     169..185
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2542..2564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2602..2620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2688..2792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2793..2861,2862..3653)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3011..3030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3824..4412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4313..4332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4539..4561
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4558..4574
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4558..4574
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    4594..4717
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4758..5063)
                     /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(4860..4879)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5383..6063)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5978..5997
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6117..6123,6124..6141,6142..6147))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(6172..6295)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6303..6320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6303..6319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6312..6334)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
       61 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
      121 ccttacgcat ctgtgcggta tttcacaccg catatgccgg gcccccctcg aggtcgacgg
      181 tatcgataag cttcagtaaa agaagtagaa ttttatagtt ttttttctgt ttgaaaaatt
      241 ctccccatca atgttctttc aaataaatac atcactaatg caaagtattc tataacctca
      301 tatctaaatt cttcaaaatc ttaacatatc ttatcattgc tttaagtcaa acgtaacatt
      361 aaaaaaaatg ttttggaaaa tgtgtcaagt ctctcaaaat tcagtttttt aaaccactcc
      421 tatagtccta tagtcctata gttacccatg aaatccttat atattactgt aaaatgtttc
      481 aaaaaccatt ggcaaattgc cagaactgaa aatttccggc aaattgggga accggcaaat
      541 tgccaatttg ctgaatttgc cggaaaaacg gtaattgccg aaagtttttg acacgaaaat
      601 ggcaaattgt ggttttaaaa tttttttttt tggaaatttc agaatttcaa ttttaatcgg
      661 caaaactgta ggcatcctaa gaatgttcct acatctattt tgaaaagtaa gcgaattcta
      721 tgaaaatgtc taaagaaaat ggggaaacaa tttcaaaaag gcacagtttc aagtgtttcc
      781 gaattatact aaatccctct aaaaacttcc ggcaaattga tatccgtaaa agagcaaatc
      841 cgcatttttg ccgaaaatta aaatttccga caaatcggca aaccggcaat ttggcgaaat
      901 ttgccggaag gcaattgccg cccacccctg ttccagaggt tcaaactggt agcaaagctc
      961 aaaatttctc aaattctcca attttttttt gaattttggc agtgtaccaa aatgacattc
     1021 agtcatattg gtttattata gatttattta gataaaatcc taaatgattc tacctttaaa
     1081 gatgcccact ttaaaagtaa tgactcaaac ttcaaattgc tctaagattc tattgaatta
     1141 ccatcttttc ctctcatttt ctctcactgt ctatttcatc acaaattcat ccctctctcc
     1201 ctctctttct ctctccctct ctctctttct ctttgctcat catctgtcat tttgtccgtt
     1261 cctctctctg cgccctcagc gttccccaca ctctctcgct tctcttttcc tagacgtctt
     1321 cttttttcat cttcttcagc cctttttcgc cattttccat ctctgtcaat cattacggac
     1381 gacccccatt aatttttgga tgtgctctgc gagcaagagc acacggaact catttatttt
     1441 ttgttggaat ttctttttca ttttagttag ttaaaagcta caacagcata tatgaaggca
     1501 gagcaacaac aacaatcgat cgcacccggc tcggcaacct tcccgtctca gatgccccga
     1561 cccccaccgg taaccgaaca ggctataacc accgaggcgg agcttctcgc gaaaaatcaa
     1621 attacaccaa atgatgtgct agctcttcca ggaatcactc aaggattcct atgctcccca
     1681 tcggcaaacg tctataacat tgagttcacc aaattccaaa tccgtgacct cgacacggaa
     1741 cacgtgctct tcgaaattgc aaaaccggaa aatgagacgg aagagaatct gcaggcgcaa
     1801 gccgaatcgg caagatatgt ccgatatcga tttgcgccga attttctgaa attaaagacg
     1861 gtcggcgcga cggtcgagtt caaggtcggc gatgtgccga tcacacattt tcgaatgatt
     1921 gaacggcact ttttcaagga tcgtttattg aaatgctttg attttgaatt tggattctgt
     1981 atgccaaatt cgagaaacaa ctgtgaacat atctatgaat ttccacaact ttcacaacaa
     2041 ctcatggacg acatgatcaa taatccaaac gagacccgct ccgatagctt ctattttgtg
     2101 gagaataagc tcgtaatgca caataaggcc gactactcgt atgatgcata atttcccgcc
     2161 gatggaataa aactaataat ttttctcgca atttatagaa atcatgtgat tttcctaccg
     2221 ctaaatttaa aatttcctaa tttttcacta tatttctcac aaaatgcatt ccgaaaactc
     2281 tcatttttca gcttttcaat acaaatccca ataaatgatt tttcaaatat aaaaaaaaaa
     2341 acggaattcc tgcagcccgg gggatccgct agttctagag cggccgccac cgcggtggag
     2401 ctcccatatg gtgcactctc agtgcaatct gctctgatgc cgcatagtta agccagcccc
     2461 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2521 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2581 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2641 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2701 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2761 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2821 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2881 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2941 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3001 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3061 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3121 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3181 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3241 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3301 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3361 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3421 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3481 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggtcag
     3541 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3601 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3661 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3721 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4441 tcctggtctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4501 tggataaccg tattaccgcc tttgagtgag ctgataccag cggataacaa tttcacacag
     4561 gaaacagcta tgaccatgat tacgccaagc tatcaacttt gtatagaaaa gttgaacgag
     4621 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     4681 cataatactg taaaacacaa catatccagt cactatggtc gacctgcaga ctggctgtgt
     4741 ataagggagc ctgacattta tattccccag aacatcaggt taatggcgtt tttgatgtca
     4801 ttttcgcggt ggctgagatc agccacttct tccccgataa cggagaccgg cacactggcc
     4861 atatcggtgg tcatcatgcg ccagctttca tccccgatat gcaccaccgg gtaaagttca
     4921 cgggggactt tatctgacag cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg
     4981 ggcgtgtcaa taatatcact ctgtacatcc acaaacagac gataacggct ctctctttta
     5041 taggtgtaaa ccttaaactg catttcacca gcccctgttc tcgtcggcaa aagagccgtt
     5101 catttcaata aaccgggcga cctcagccat cccttcctga ttttccgctt tccagcgttc
     5161 ggcacgcaga cgacgggctt cattctgcat ggttgtgctt accgaaccgg agatattgac
     5221 atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa ctgtcactgt aatacgctgc
     5281 ttcatagcat acctcttttt gacatacttc gggtatacat atcagtatat attcttatac
     5341 cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc ttttagtaag ccggatcctc
     5401 tagattacgc cccgcctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     5461 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     5521 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     5581 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     5641 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     5701 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     5761 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     5821 ctcaccgtct ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat
     5881 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     5941 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     6001 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     6061 cattttagct tccttagctc ctgaaaatct cgacggatcc taactcaaaa tccacacatt
     6121 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgcggccg ccatagtgac
     6181 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     6241 atattgatat ttatatcatt ttacgtttct cgttcaactt tattatacat agttgataat
     6301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaa
//
