LOCUS       sequence-46085-e        4121 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 4121)
  TITLE     NLdDIS
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4121)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCR4-CTE
FEATURES             Location/Qualifiers
     source          1..4121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(493..512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(494..512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(519..536)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(519..535)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(528..550)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             673..975
                     /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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     854..873
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1324..2118
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1324..2118
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1378..1397)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1988..2007
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(2368..3159,3160..3228))
                     /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     2991..3010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3352..3940
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3841..3860
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4094..4111
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc ccttgggccc
      301 acacatccct cggaggctgc gcctgtctta ggttggagtg atacatttct tgtcactatt
      361 acccgtcatt ggattaggca gtagctttga cggccctcct gtcttaggtt agtgagaaat
      421 gtcactctct tacccgtcat tggctgtcca gtcttctaga agggcgaatt cgcggccgct
      481 aaattcaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
      541 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      601 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      661 ctatacgtac ggcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
      721 gtggatgtac agagtgatat tattgacacg ccggggcgac ggatggtgat ccccctggcc
      781 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
      841 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
      901 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
      961 ttctggggaa tataaatgtc aggcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1021 ttcacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     1081 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     1141 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     1201 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     1261 ccaaggatct gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt
     1321 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     1381 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     1441 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     1501 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     1561 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     1621 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     1681 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     1741 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     1801 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     1861 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     1921 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     1981 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2041 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2101 cttgacgagt tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt
     2161 acgcatctgt gcggtatttc acaccgcatc aggtggcact tttcggggaa atgtgcgcgg
     2221 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagattatc
     2281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     2821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     2881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     2941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3241 tcaatattat tgaagcattt atcagggtta ttgtctcatg accaaaatcc cttaacgtga
     3301 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3361 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3421 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3481 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3541 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3601 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3661 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3721 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3781 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3841 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3901 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3961 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4021 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4081 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
