LOCUS       sequence-10403-e        4099 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 4099)
  TITLE     Hudson lab plasmid
REFERENCE   2  (bases 1 to 4099)
  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: pCR-Myt1NB580
FEATURES             Location/Qualifiers
     source          1..4099
                     /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        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(986..1005)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(987..1005)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1012..1029)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1012..1028)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1021..1043)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1166..1468
                     /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     1347..1366
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             1817..2611
                     /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             1817..2611
                     /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(1871..1890)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2481..2500
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      2812..2821
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2818..3192
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3330..3918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3819..3838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4072..4089
                     /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 gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
      301 ctagtaacgg ccgccagtgt gctggaattc gcccttcagc ttgcttgatc gtttccagaa
      361 ggcctttgtt ctccggattc tggtagcagt cctgattgga gtacatgtca gtgagggtgt
      421 tcacataggc gtcgaaattc tgttcacata ttggttccat gtgtggaagg cgaatattgg
      481 caagactttg gatgagggct tggctaagcc cggacaattc cagaaacagg gcctcattct
      541 gctcctcaat gagcttgttc tcctcctcga tgttcttcag gttcctctcc atggacgaga
      601 tctgagactg cagctgcacc atggcagcct ccattccgga attggactca tttaagtctc
      661 gtatctcctg atttagctgc ttgatctcct catcattctc cagcacatca cttgtcttga
      721 actttgggct gagaatctca tcccctgaga gttttccttt ctttccagca aaggttgctc
      781 tgggacagcc agacaaactt cggtgggtga ggaaactgcc gttggcatgg ccagagccgt
      841 cgcaacctgg ggtggggcaa gtagggccct cattcttcag cgacttccac gagaaagatg
      901 aaccattgag agctccaagg gcgaattctg cagatatcca tcacactggc ggccgctcga
      961 gcatgcatct agagggccca attcgcccta tagtgagtcg tattacaatt cactggccgt
     1021 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1081 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1141 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
     1201 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
     1261 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     1321 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     1381 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     1441 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt
     1501 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     1561 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     1621 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     1681 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     1741 ggctttctcg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     1801 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     1861 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     1921 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     1981 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2041 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2101 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2161 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2221 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2281 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2341 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2401 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2461 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2521 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     2581 cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt tcctgatgcg
     2641 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacaggtgg cacttttcgg
     2701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2761 ctcatgagac aataaccctg ataaatgctt caataatagc acgtgaggag ggccaccatg
     2821 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     2881 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     2941 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     3001 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     3061 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     3121 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     3181 gagcaggact gacacgtgct aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct gggcttttgc
     3961 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4021 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4081 gtgagcgagg aagcggaag
//
