LOCUS       sequence-83400-e        4983 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 4983)
  AUTHORS   Hartzog PE, Nicholson BP, McCusker JH
  TITLE     Cytosine deaminase MX cassettes as positive/negative selectable 
            markers in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2005 Jul 30;22(10):789-98.
  PUBMED    16088873
REFERENCE   2  (bases 1 to 4983)
  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: "Yeast."; 
            date: "2005-07-30"; volume: "22(10)"; pages: "789-98"
COMMENT     Sequence Label: pBN27
FEATURES             Location/Qualifiers
     source          1..4983
                     /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"
     primer_bind     complement(1439..1458)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1440..1458)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1465..1482)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1465..1481)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1474..1496)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1623..2003
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1710..1729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1920..1941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2371..3165
                     /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
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2425..2444)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             join(3183..3251,3252..4043)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4214..4802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4703..4722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4956..4973
                     /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 gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttcggaat
      301 ggtcagggtg gtacaatttt acataacgat gatatttttt ttttaacgac ttcttgtcca
      361 gtttaggatt tccagatccg gcctttggaa tgccaaaaat atcataggga gttggatctg
      421 ccaactcagg ccattgttca tcccttatcg taagttttct attgccattt ttatcgttcg
      481 ctgtagcata cttagctata aaagtgattt gtgggggaca cttttctaca catgataagt
      541 gccacttgaa taaaaatggg tatacgaact tatggtgtag cataacaaat atattgcaag
      601 tagtgaccta tggtgtgtag atatacgtac agttagttac gagcctaaag acacaacgtg
      661 tttgttaatt atactgtcgc tgtaatatct tctcttccat tatcaccggt cattccttgc
      721 aggggcggta gtacccggag accctgaact tttctttttt tttttgcgaa attaaaaagt
      781 tcattttcaa ttcgacaatg agatctacaa gccattgttt tatgttgatg agagccagct
      841 taaagagctg aacctgcaga caagtccagc acgctgtatt tacgtattta attttatata
      901 tttgtgcata cactactagg gaagacttga aaaaaaccta ggaaatgaaa aaacgacaca
      961 ggaagtcccg tatttactat tttttccttc cttttgatgg ggcagggcgg aaatagagga
     1021 taggataagc ctactgctta gctgtttccg tctctacttc ggtagttgtc tcaattgtcg
     1081 tttcagtatt acctttagag ccgctagacg atggttgagc tatttgttga gggaaaacta
     1141 agttcatgta acacacgcat aacccgatta aactcatgaa tagcttgatt gcaggaggct
     1201 ggtccattgg agatggtgcc ttattttcct tataggcaac gatgatgtct tcgtcggtgt
     1261 tcaggtagta gtgtacactc tgaatcaggg agaaccaggc aatgaacttg ttcctcaaga
     1321 aaatagcggc cataggcatg gattggttaa ccacaccaga tatgcttgga agggcgaatt
     1381 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc
     1441 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     1501 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     1561 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     1621 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     1681 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     1741 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     1801 tagagcttta cggcacctcg accgcaaaaa acttgatttg ggtgatggtt cacgtagtgg
     1861 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     1921 tggactcttg ttccaaactg gaacaacact caaccctatc gcggtctatt cttttgattt
     1981 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaattca
     2041 gggcgcaagg gctgctaaag gaaccggaac acgtagaaag ccagtccgca gaaacggtgc
     2101 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     2161 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
     2221 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     2281 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct
     2341 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2401 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2461 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2521 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     2581 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2641 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcgcct tgctcctgcc
     2701 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     2761 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     2821 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     2881 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgat ccatggcgat
     2941 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcaa cgactgtggc
     3001 cggctgggtg tggcggaccg ctatcaggac atagcgttgg atacccgtga tattgctgaa
     3061 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3121 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattga aaaaggaaga
     3181 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3241 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3301 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3361 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtcat acactattat
     3421 cccgtattga cgccgggcaa gagcaactcg gtcgccgggc gcggtattct cagaatgact
     3481 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3541 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3601 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3661 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagagt gacaccacga
     3721 tgcctgtagc aatgccaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3781 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3841 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3901 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3961 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4021 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4081 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4141 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4201 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4261 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4321 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     4381 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4441 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4501 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4561 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     4621 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     4681 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4741 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4801 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4861 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4921 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4981 aag
//
