LOCUS       sequence-38146-e        7980 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 7980)
  AUTHORS   Meissner B, Rogalski T, Viveiros R, Warner A, Plastino L, Lorch A, 
            Granger L, Segalat L, Moerman DG
  TITLE     Determining the Sub-Cellular Localization of Proteins within 
            Caenorhabditis elegans Body Wall Muscle.
  JOURNAL   PLoS One. 2011  . 6(5):e19937.
  PUBMED    21611156
REFERENCE   2  (bases 1 to 7980)
  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: "PLoS One. 
            2011  . 6(5):e19937."
COMMENT     Sequence Label: pDM#834
FEATURES             Location/Qualifiers
     source          1..7980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1893..2017
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2042..2047,2048..2065,2066..2072)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2126..2785
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2192..2211)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3127..3432
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3311..3330
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3473..3597)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3650..3678)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4426..4446
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5420..5442)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5542..5561
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5639..5657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5725..5829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5830..5898,5899..6690)
                     /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(6048..6067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6861..7449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7350..7369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7603..7620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7737..7758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7773..7778,7779..7796,7797..7803)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7811..7827
                     /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     7816..7838
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7835..7851
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7835..7851
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttttcc tatctccggc ttgccttcac attcgcaaac aaagcctgag aatttgatga
       61 gtatttgttc catttgttgc atcaaccagc atgttgtata actgttcatt ttacgattcc
      121 aataagtttt tatgatggag tactgttaat agcgttttta ctttaaaatt ctcaaactgg
      181 ttagcagacc aattttgggt ctgctaaata gtttttataa aaaaaccatt gacagtacag
      241 cctctaaaaa agattaaaat aatttttgaa ttcagaaagt tcttgtttca tataaataag
      301 agccatccct ctacacgggt catctagtag ctctaggtcg acacctgttc gtagcaggtt
      361 ttatgagctc aacaaaggtt gtaggtgagc cgtgccaacg tctcagtttc aaattccgag
      421 ttgagtatta cgaaaacctg ggtataaaac gaggggatgc cgagaaaaaa agagtagtgg
      481 acacacacac acatacgtct catcttctta tttcttttcg tttcttcccg tccaggagcc
      541 acttgtcgtc ccgtcgtcat tttgagttcc aaatgatgaa agattcaatg aatcacggag
      601 acgcagggtc ccgtcgatgg ctcattttgc gtgttttagt ggcgggacac aggaagaagt
      661 cgaagattca caacatgtcg caactgattc atcactagtg ccttcttgct ccaagagcaa
      721 tgtttttcta attagtttgt actacttgat aacttcttaa gatttgttct actactttta
      781 aaaatttggt tttgtagcta gaaatcctga aaaaaaaaac atttttgcta tgaacttttt
      841 tttggatata cttttataaa cagcatctag aattagttag agtacctgaa aaatcaaaaa
      901 tacgaaatag tacttgtatc ctatgtgcga agttgcacga ttttcattgc aaaattaggg
      961 tctccggtct cgcgacgacc gaagaaaata taaataaatt gaaatgaaaa cgagtgtgcg
     1021 cctttaagaa gtacagtagt tttttttgaa agtttccttg aatttttttt ccacttcaaa
     1081 ttattcgttt cgcttcgaga ccaagtaccg taatagtacc gtaaatttcg aattcctatt
     1141 ttttaaaaaa attctttaaa ctgaatttaa tcaaaaatca atattcaagt taaattttca
     1201 cctttttggt gagtttcaat ttctcgaatg atacagtaat tttgaaaaaa taattttttc
     1261 ttatttcaaa taatctaaaa aataaaactt catattcttt caatgatgcc gaaatttaca
     1321 aaagccatgg tagtttatta gaatttttta gaatttcatg tcaaaaaatt ggaatagttt
     1381 tcacctatca taatttggaa tttgtaattt tgaccaaaaa agtcctacga attttgtatt
     1441 taaattcaag aatcatagaa cttcgaacct gcaggaatgt ccaaaattgt tagactttaa
     1501 ttttttgttg gggaattgtt ggtttttttt tttgaaaaag tgttagacct atatttgtat
     1561 gagtacaaat gctttaaaca taattagaaa caaaaacaat ttaaaaaaat ctttaattat
     1621 ttggttgact tccaaaattg ttgagaactt gcaatttttg gatatagatg gaaccaattt
     1681 caattttttt aaataaaaaa gtattgctat gatatttgac aattcatctt cttctctgta
     1741 ttttccgtac cccaatggtt tgcacttcaa tataagtttt aaatttaaaa aagttgattt
     1801 ccttgtttcc aaaacacttc tatgccttaa aatgtgataa aaatataata aacgattcaa
     1861 tattaattca gaaaatgccc cgggattggg gcacaagttt gtacaaaaaa gctgaacgag
     1921 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     1981 cataatactg taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg
     2041 ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccggcgag
     2101 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     2161 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     2221 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     2281 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     2341 attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     2401 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     2461 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     2521 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     2581 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     2641 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     2701 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     2761 cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca
     2821 gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg
     2881 aagtatgtca aaaagaggtg tgctatgaag cagcgtatta cagtgacagt tgacagcgac
     2941 agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca
     3001 tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga
     3061 tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag aacagggact
     3121 ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
     3181 gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt
     3241 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     3301 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     3361 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     3421 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga
     3481 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     3541 tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgag
     3601 gggggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag ttgtcccaat
     3661 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     3721 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     3781 tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt taaattttca
     3841 gccaacactt gtcactactt tctgttatgg tgttcaatgc ttctcgagat acccagatca
     3901 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     3961 tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt tcggtactaa
     4021 ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg atacccttgt
     4081 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     4141 attggaatac aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg
     4201 aatcaaagtt gtaagtttaa acatgatttt actaactaac taatctgatt taaattttca
     4261 gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     4321 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     4381 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     4441 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagc attcgtagaa
     4501 ttccaactga gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa ataatagggg
     4561 ccgctgtcat cagagtaagt ttaaactgag ttctactaac taacgagtaa tatttaaatt
     4621 ttcagcatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca acatccctac
     4681 atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa aacttcttct
     4741 taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt gtgtagattc
     4801 aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa
     4861 taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg ttttttttac
     4921 ttctgataaa ttttttttga aacatcatag aaaaaaccgc acacaaaata ccttatcata
     4981 tgttacgttt cagtttatga ccgcaatttt tatttcttcg cacgtctggg cctctcatga
     5041 cgtcaaatca tgctcatcgt gaaaaagttt tggagtattt ttggaatttt tcaatcaagt
     5101 gaaagtttat gaaattaatt ttcctgcttt tgctttttgg gggtttcccc tattgtttgt
     5161 caagagtttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     5221 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     5281 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     5341 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     5401 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5461 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5521 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5581 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     5641 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5701 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5761 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5821 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5881 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5941 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6001 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6061 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6121 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6181 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6241 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6301 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6361 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6421 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6481 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6541 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6601 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6661 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6721 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6781 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6841 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6901 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6961 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     7021 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7081 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7141 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7201 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     7261 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7321 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7381 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7441 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7501 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7561 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7621 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7681 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7741 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7801 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7861 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     7921 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
