LOCUS       sequence-495443-        7834 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7834)
  AUTHORS   Larson DR, Zenklusen D, Wu B, Chao JA, Singer RH
  TITLE     Real-time observation of transcription initiation and elongation on 
            an endogenous yeast gene.
  JOURNAL   Science. 2011 Apr 22;332(6028):475-8.
  PUBMED    21512033
REFERENCE   2  (bases 1 to 7834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2011-04-22"; volume: "332(6028)"; pages: "475-8"
COMMENT     Sequence Label: pDZ306 (GLT1 POL1 24xPP7SL)
FEATURES             Location/Qualifiers
     source          1..7834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        421..764
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             765..1463
                     /codon_start=1
                     /product="S. kluyveri imidazoleglycerol phosphate 
                     dehydratase"
                     /label=k-HIS3
                     /translation="MAEPAQKKQKQTVQERKAFISRITNETKIQIAISLNGGYIQIKDS
                     ILPAKKDDDVASQATQSQVIDIHTGVGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHT
                     TEDCGIALGQAFKEAMGAVRGVKRFGTGFAPLDEALSRAVVDLSSRPFAVIDLGLKREM
                     IGDLSTEMIPHFLESFAEAARITLHVDCLRGFNDHHRSESAFKALAVAIREAISSNGTN
                     DVPSTKGVLM"
     terminator      1467..1664
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3912..3931)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3913..3931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3938..3955)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3938..3954)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3969)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             4092..4394
                     /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     4273..4292
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             4743..5537
                     /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             4743..5537
                     /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(4797..4816)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5407..5426
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(5787..6578,6579..6647))
                     /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     6410..6429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6771..7359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7260..7279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7513..7530
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7647..7668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7683..7688,7689..7706,7707..7713)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7721..7737
                     /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     7726..7748
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7745..7761
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7745..7761
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7782..7800
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atatattaat taaaccgttc aaccctatac agcgcaaatt tggtcctaat acacttcggt
       61 tttaatgcgt caatccgatt ggctccgata agcttttgca catttttcaa gtcatatgtc
      121 acgacgaacg tgatgcccgc aaaacgtaaa aaaaagaagc aataaattgc ccaaaaaaaa
      181 aatataaact gcttttctta gtcattcata agttctgatg gtcttcctta tcttattgta
      241 tttttcgtcc tattgtttca tttcttacca aattaatcaa ttcttatatc ttacttgata
      301 acacaccaaa ctaatcgtct ccacatcata ggaagatagg aaattgctat ctcagtccta
      361 tactacgcag acctcgagtc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
      421 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
      481 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      541 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      601 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      661 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      721 ttgctaggat acagttctca catcacatcc gaacataaac aaccatggca gaaccagccc
      781 aaaaaaagca aaaacaaact gttcaggagc gcaaggcgtt tatctcccgt atcactaatg
      841 aaactaaaat tcaaatcgct atttcgctga atggtggtta tattcaaata aaagattcga
      901 ttcttcctgc aaagaaggat gacgatgtag cttcccaagc tactcagtca caggtcatcg
      961 atattcacac aggtgttggc tttttggatc atatgatcca tgcgttggca aaacactctg
     1021 gttggtctct tattgttgaa tgtattggtg acctgcacat tgacgatcac catactaccg
     1081 aagattgcgg tatcgcatta gggcaagcgt tcaaagaagc aatgggtgct gtccgtggtg
     1141 taaaaagatt cggtactggg ttcgcaccat tggatgaggc gctatcacgt gccgtagtcg
     1201 atttatctag tagaccattt gctgtaatcg accttggatt gaagagagag atgattggtg
     1261 atttatccac tgaaatgatt ccacactttt tggaaagttt cgcggaggcg gccagaatta
     1321 ctttgcatgt tgattgtctg agaggtttca acgatcacca cagaagtgag agtgcgttca
     1381 aggctttggc tgttgccata agagaagcta tttctagcaa tggcaccaat gacgttccct
     1441 caaccaaagg tgttttgatg tgaagtactg acaataaaaa gattcttgtt ttcaagaact
     1501 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     1561 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     1621 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     1681 aacgccgcca tccagagctc tacatgaact gaccgaaatt gcagcatcct tacaaaaacg
     1741 gatacatacg ttagaaacgg aaaacaagct tttaaagaat ttagttctga gtagcggtga
     1801 aacggaagga ataaaaaaag ctgaaagctt aaagaagcaa atttttgaga aggttcagaa
     1861 agaataaaat aacagactat acagtttaat tatatatata tatatatata cgtgcatttt
     1921 tcttaaagaa atataacttt tttcttgtac tgcctgcaat ctctattctt cattcatcac
     1981 acatctattc aaacgcgtta aaattttttt gttagcctat aagtaaaatg cgtcgcgtgt
     2041 atccaatttt actgaattaa attctttact cattggatga tatatatctt cttggtctag
     2101 acaacgaaca cataccaatt cagactcacg agaaaaaaaa ctgtcttcac tctgctatta
     2161 gaaaccaaat agtatcaagg cataaaataa acagtggaaa ggatcctaag gtacctaatt
     2221 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca
     2281 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat
     2341 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc
     2401 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa
     2461 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag
     2521 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta
     2581 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca
     2641 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct
     2701 aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc
     2761 agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc
     2821 taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac
     2881 cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac
     2941 ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa
     3001 ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta
     3061 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     3121 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt
     3181 acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga
     3241 aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg
     3301 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     3361 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag
     3421 gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta
     3481 gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatccatg
     3541 tcccaggata gaattttgtt agatagatct atgccagtgt tgaaatcaga caatttcgat
     3601 ccattggaag aagcttacga aggtgggaca attcaaaact ataacgatga acaccatctt
     3661 cataaatctt gggcaaatgt gattccggac aaacgaggac tttacgaccc tgattatgaa
     3721 catgacgctt gtggtgtcgg tttcgtagca aataagcatg gtgaacagtc tcacaagatt
     3781 gttactgacg ctagatatct tttagtgaat atgacacatc gtggtgccgt ctcatctgat
     3841 gggaacggtg acggtgccgg tattctgcta ggtatcccaa gggcgaattc gcggccgcta
     3901 aattcaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     3961 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4021 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4081 tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     4141 tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca
     4201 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     4261 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     4321 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     4381 tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt
     4441 tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     4501 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     4561 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     4621 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     4681 caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc
     4741 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4801 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4861 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4921 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4981 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5041 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5101 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5161 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5221 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5281 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5341 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5401 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5461 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5521 ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta
     5581 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagattatca
     5701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5821 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5881 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5941 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6001 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6061 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6121 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6181 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6241 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6301 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6361 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6421 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6481 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6541 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6601 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6661 caatattatt gaagcattta tcagggttat tgtctcatga ccaaaatccc ttaacgtgag
     6721 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6781 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6841 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6901 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6961 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7021 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7081 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7141 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7201 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7261 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7321 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7381 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7441 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7501 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7561 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7621 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7681 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7741 cacacaggaa acagctatga ccatgattac gccaagctca gaattaaccc tcactaaagg
     7801 gactagtcct gcaggtttaa acgaattcgc cctt
//
