LOCUS       sequence-39-expo        5808 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 5808)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 5808)
  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: pDD16_01
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             163..183
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             271..3318
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(273..295)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3382..3404)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3504..3523
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3601..3619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5565..5582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5735..5740,5741..5758,5759..5765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5789
                     /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     5778..5800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5797..5808,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5797..5808,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggat
       61 tggccaaagg acccaaaggt atgtttcgaa tgatactaac ataacataga acattttcag
      121 gaggaccctt ggagggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      181 gtaccggtgg gtgaagacca gaaacagcac ctcgaactga gccgcgatat tgcccagcgt
      241 ttcaacgcgc tgtatggcga gatcgatccc gtcgttttac aacgtcgtga ctgggaaaac
      301 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      361 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      421 cgctttgcct ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt
      481 cctgaggccg atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc
      541 atctacacca acgtgaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat
      601 ccgacgggtt gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag
      661 acgcgaatta tttttgatgg cgttaactcg gcgtttcatc tgtggtgcaa cgggcgctgg
      721 gtcggttacg gccaggacag tcgtttgccg tctgaatttg acctgagcgc atttttacgc
      781 gccggagaaa accgcctcgc ggtgatggtg ctgcgctgga gtgacggcag ttatctggaa
      841 gatcaggata tgtggcggat gagcggcatt ttccgtgacg tctcgttgct gcataaaccg
      901 actacacaaa tcagcgattt ccatgttgcc actcgcttta atgatgattt cagccgcgct
      961 gtactggagg ctgaagttca gatgtgcggc gagttgcgtg actacctacg ggtaacagtt
     1021 tctttatggc agggtgaaac gcaggtcgcc agcggcaccg cgcctttcgg cggtgaaatt
     1081 atcgatgagc gtggtggtta tgccgatcgc gtcacactac gtctgaacgt cgaaaacccg
     1141 aaactgtgga gcgccgaaat cccgaatctc tatcgtgcgg tggttgaact gcacaccgcc
     1201 gacggcacgc tgattgaagc agaagcctgc gatgtcggtt tccgcgaggt gcggattgaa
     1261 aatggtctgc tgctgctgaa cggcaagccg ttgctgattc gaggcgttaa ccgtcacgag
     1321 catcatcctc tgcatggtca ggtcatggat gagcagacga tggtgcagga tatcctgctg
     1381 atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt atccgaacca tccgctgtgg
     1441 tacacgctgt gcgaccgcta cggcctgtat gtggtggatg aagccaatat tgaaacccac
     1501 ggcatggtgc caatgaatcg tctgaccgat gatccgcgct ggctaccggc gatgagcgaa
     1561 cgcgtaacgc gaatggtgca gcgcgatcgt aatcacccga gtgtgatcat ctggtcgctg
     1621 gggaatgaat caggccacgg cgctaatcac gacgcgctgt atcgctggat caaatctgtc
     1681 gatccttccc gcccggtgca gtatgaaggc ggcggagccg acaccacggc caccgatatt
     1741 atttgcccga tgtacgcgcg cgtggatgaa gaccagccct tcccggctgt gccgaaatgg
     1801 tccatcaaaa aatggctttc gctacctgga gagacgcgcc cgctgatcct ttgcgaatac
     1861 gcccacgcga tgggtaacag tcttggcggt ttcgctaaat actggcaggc gtttcgtcag
     1921 tatccccgtt tacagggcgg cttcgtctgg gactgggtgg atcagtcgct gattaaatat
     1981 gatgaaaacg gcaacccgtg gtcggcttac ggcggtgatt ttggcgatac gccgaacgat
     2041 cgccagttct gtatgaacgg tctggtcttt gccgaccgca cgccgcatcc agcgctgacg
     2101 gaagcaaaac accagcagca gtttttccag ttccgtttat ccgggcaaac catcgaagtg
     2161 accagcgaat acctgttccg tcatagcgat aacgagctcc tgcactggat ggtggcgctg
     2221 gatggtaagc cgctggcaag cggtgaagtg cctctggatg tcgctccaca aggtaaacag
     2281 ttgattgaac tgcctgaact accgcagccg gagagcgccg ggcaactctg gctcacagta
     2341 cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg ggcacatcag cgcctggcag
     2401 cagtggcgtc tggcggaaaa cctcagtgtg acgctccccg ccgcgtccca cgccatcccg
     2461 catctgacca ccagcgaaat ggatttttgc atcgagctgg gtaataagcg ttggcaattt
     2521 aaccgccagt caggctttct ttcacagatg tggattggcg ataaaaaaca actgctgacg
     2581 ccgctgcgcg atcagttcac ccgtgcaccg ctggataacg acattggcgt aagtgaagcg
     2641 acccgcattg accctaacgc ctgggtcgaa cgctggaagg cggcgggcca ttaccaggcc
     2701 gaagcagcgt tgttgcagtg cacggcagat acacttgctg atgcggtgct gattacgacc
     2761 gctcacgcgt ggcagcatca ggggaaaacc ttatttatca gccggaaaac ctaccggatt
     2821 gatggtagtg gtcaaatggc gattaccgtt gatgttgaag tggcgagcga tacaccgcat
     2881 ccggcgcgga ttggcctgaa ctgccagctg gcgcaggtag cagagcgggt aaactggctc
     2941 ggattagggc cgcaagaaaa ctatcccgac cgccttactg ccgcctgttt tgaccgctgg
     3001 gatctgccat tgtcagacat gtataccccg tacgtcttcc cgagcgaaaa cggtctgcgc
     3061 tgcgggacgc gcgaattgaa ttatggccca caccagtggc gcggcgactt ccagttcaac
     3121 atcagccgct acagtcaaca gcaactgatg gaaaccagcc atcgccatct gctgcacgcg
     3181 gaagaaggca catggctgaa tatcgacggt ttccatatgg ggattggtgg cgacgactcc
     3241 tggagcccgt cagtatcggc ggaattccaa ctgagcgccg gtcgctacca ttaccaactt
     3301 gtctggtgtc aaaaataata ggcctactag tcggccgtac gggccctttc gtctcgcgcg
     3361 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3421 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3481 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3541 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg
     3601 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagct
//
