LOCUS       sequence-11288-e        9537 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 9537)
  TITLE     George Cross Lab Plasmids
REFERENCE   2  (bases 1 to 9537)
  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: pT13-11 episome
FEATURES             Location/Qualifiers
     source          1..9537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             5541..6335
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5595..5614)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6205..6224
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(6732..6750)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6733..6750)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7041..7060)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7271..7290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7390..7412
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7450..7468)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7536..7640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7641..7709,7710..8501)
                     /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(7859..7878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8672..9260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9161..9180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9414..9431
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     9512..9531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9512..9530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaattcgcgc tctctaattg cttgttcttt ggcaatgtcc aaaatgatac tgttaagctc
       61 ctccatgatt tcaatagcag ttggggaatt gtatcttcca tctcgcttca actctagaag
      121 ttccctttct tttcccaaga ataattcact atcgtcaacg ggaagctcgt ctagagtaaa
      181 gccgagtggg tgttgctcga gagataacct cacatcactt cgcagcttac gtgctatgtc
      241 ctcagaacgt ctccgcatcg cccctgcaag gtcctggatt ttcaaagtat ccttagcatc
      301 aaaaacagct ttaatgtatc cattctccaa ctcaataaac ggtaagtcat tgtcaagagg
      361 aagcaccgac aacggaagac catcaggtga ggtgtcaaga acatcctctc tcatttttcg
      421 cttcacctca agggctaaat cattcacgta gtccctcatg gcattttcca cgcccaccaa
      481 cttatgatca ttctcgcgaa tgttcagacc cttcaatgtc aagcgctcca cctctagcaa
      541 atgaaagtta tcatcatcat caagaggtac acaatgcaat gggatgccat gaggctcagg
      601 atcgagaaac gacctctcac tacgcaactt taacaacgca agcttttccg ctcgcccatt
      661 cagggcnttc tccacactgg caattgagtc cttgttaccc accacgtcac gcattagcgc
      721 agctcgtttc acttcaagca tcgaaaattc atcgtcattg tccagtggca catcatccaa
      781 aggtacacca caagggtgag aaatcaggta atccttccgc tccgcacagt tcttcatttt
      841 agcaagctga tgcgcacgac catccagcaa ctcctcaatg gcagcaattt tctgtgcctc
      901 aggaggagac tccttacaaa gaagccgtag ttgcttttca agagaacgaa actccttatc
      961 gctatctaga ggcaattcct ccagtagacg accctcaggc tcagggaaga gaaaagcacg
     1021 atctttagaa atcatctccg ccgccatctg cttggctaac tcgttcaagt cagacttcag
     1081 caaagcaaca tcggcaccat ttgcccgcag gtccttccga agcttgtgca gttcagcttc
     1141 tttcgcaaca aaggtctcgt tattgtcaag gggcagctca tctagggaca cccccaatgg
     1201 catcgggtcc agaaacatgc ggatctcaca acgcatctta tcagccagtt cgatcaccct
     1261 ctcattcatc gcatcttcaa gatttcgtat cgctgctata ttagcatgcc ggtcctcgca
     1321 caaaccaaac cgacgaattt caagatcaca aaaaacctcg tcgttatcaa taggaacatc
     1381 ctcaatagca ataccaccag ggttagggtc cataaatttc gatcgctccg cgtgcttcat
     1441 ttccgtagca agaacgtgtg cacgctcgtt caggccaccc tccaaatccg agatggcacg
     1501 tccgttcctc tgagggttct tcttcagtag cgcacgttta atttcaagct cctggaacac
     1561 actatcgcta tcaagtggta gcaaacgaag cggcacacca acagatgctg ggtccaagta
     1621 accacggtca cttcgacaag tgctccctcg caagggcttc cgcacgatca ttcagtgcac
     1681 gctccacatc cattattact gtgtcctgtt tcttcgcatc acacttgaga cgttgccgat
     1741 gcacctcaag ctcgtggaat tcagagcgaa tttcttattg aaaagaaaaa ttgtatcgtt
     1801 ttaagagtta ttatcatttg caaccagctt cctttttgca ggcatcttcc aatttacctg
     1861 aatatttctt ttccctcttc tcatctttcc ctcttttttc ttggttttct ttttctattt
     1921 tttctttcac agaatttttg aacttgcatt tgttggtgtt gaaactgcac cctttatcat
     1981 ctactccctt atatttgact ttgttggtgt ctgctgaatt gcattccttt tccttgtttt
     2041 ctgtttgttt tttccttttg tcatcgcctc ttgttggttg caaattggat tcctcgacct
     2101 gttgtagatc tgccagattt tcttgtgatg caacccttag tagcatcttg aggttagcga
     2161 gtatttgggc gtcatgtgcc acgtttgctg cgtgctgctc tatgagtctt ttcgctgctg
     2221 ataactgtag catccacggt atattgtatg gcattgagtc tgctacggat ttgtaagtga
     2281 tgcagcctgc gccgttgctg ctgtcgcagt tattgcctga ttctggtgtg ccgtaaagca
     2341 ccgctttgtg ctccgcttcc tcggtgttgt tgccgagtaa atctgtgaat tggtgctgtg
     2401 ctgccgttag gtttgcagcc tataatactt caattcgcag cggtttgcat gtcgctgcta
     2461 gtcctgcgta tttttgagcc gctgctgccg cgttactgtg ggtgatagtt gccaagccgc
     2521 cgaatcctat gcaacgcttt tcataggcat ttgtcgagtc gtggctacag aggcagaccg
     2581 tatcactgcc taatcttttg cctgtacctc ccaaggtagc gccgtcggct tcttatgctg
     2641 ctccagcgcc attagcactg aatgagtctg tttctgcttc tacttttact tcttttttgc
     2701 ctgtggttac agccgnnaan tctctttggt atcatgcaca tgatgctgaa catagtcctc
     2761 cgatgcgacg gtttcgagga cacacacaag agagtgccaa gctcaatgtc agttgggaag
     2821 ttgaaggccc ttgtgcgtgc agtgtatagc gtggctccgt cccatcagca tttgtccttc
     2881 gtgtcaggtg atgctggagt ggtagaggca ccgacacaac tggataatga gctgcagagc
     2941 ctttcattct acggcatgtg tgaccgggcc atagtactgg tagttgacac atcccttcgg
     3001 agttgatgca tgagcctcgc aacactccct gaagcgaagc gaagagaatt gaaatattaa
     3061 ttcgcagccg tgccaccgct gcaaagcaac caaccttatg cagctctcgt tgttcttata
     3121 tgattgatta ctctccgttt gtgaaacctt atgcgcggtg gttccctgga gagccctgat
     3181 gatgacgcca actgagggtc ttttctagcg gccgcttttc gtagggaaca tgcacacggt
     3241 gaggggtgat ttaagttgga aatacgaatg gtagcggata ttctgtttta ctccacatag
     3301 atatgaggcc gcgcgtataa ataaataaaa ataaatgggc gtatctctcg ctaaccttca
     3361 acacaaacaa tatcggtccg acttccttcc taatattcgt cgcgcaacct tncggttttc
     3421 tttttttttt ttcacgtctt atttagaact cgcttgaaca cagggacttg aataacttca
     3481 ttagcccaac ggcggagatc cacgtgagtg atttcatgca gagactcttt tttagatcga
     3541 catgcgcacg tgtcagatgt gccgacctgc ggctcaaacc accggtatcc atcccgcaag
     3601 tgactttcca aactgtcgct ttttccagca gcgcagcggg tgctgccgtg aggagcggag
     3661 ccgaagggac gaatacaaca acaaaacgtc gcagtggttt cggcggcggt gccggtggag
     3721 gcgggcccgt gaaggatcgc tatccgcccc cagcaactga tctcggtgat ccgcggcgac
     3781 gcctggccgt gcttgtggat gcctcaagtg ttgacccccg cgtgtttcgc aacacaattc
     3841 tgccagcagc gctgaaggtt ggcacgccag tactcatacg tatattttcg gtgcagctga
     3901 gcagtgaatg ggaacctctc gtggccggca gcggagggac aagggaatca gcaacctcaa
     3961 cggaaccgga gtggaagacg tctggcaccg ctaatgatgg gagggaggaa gttagtgggg
     4021 ccgcggggtc tcgcgacagc ggcgcgtggt gtccgccggt ggagtttttc agggtggagc
     4081 gcttcattcc tgtgncaatg caaatgnagn ccgatgcana tcatatattt gncttccgta
     4141 nacaanacca gattnagncg gtggngtttg tgtgcgncna ggtagatcnc ggcgtgtncg
     4201 aaggattcct tcctaatata gcaggtaatg ggttcaatca atatgtgtta gatgagcttg
     4261 gtatggcgaa ggaggtgctt gaagatggaa gatccgaaga cgggacgccc acagtgcaga
     4321 aataaaaata atgggtgcaa gaaggcagat aatgtgaggg ttgatggagg agagggctat
     4381 ttaagaaggt gaaaggagtt tagagtacag caagggaacc ggtgccaagt attcctgtgg
     4441 aggaaaggtg tacctttttg cagggtttct cattcccaga atgcacacac gcacaccctn
     4501 tgccntttcg tattgtngnc agtggcgttc ggtaaacgcc accgccgagc tgtacatgcg
     4561 ggaaaggggc agagggttca ttgtacaatg agtcggcgtt actttctccc tttatccttc
     4621 actgcttgta ataaaaccct ctttttgctt tctcatagtc gtgttcatcc ttatttcccc
     4681 actttccgct gttctctgtt cacttatgtg cgtgcacata cgtacattat gtgttgtatt
     4741 ttgcgcaacg cccacggtac tagtaaatgg gatttgtcga ccgtggtggc ggtcgtgaag
     4801 cgaaggggtc gagcctacgg cactccaaga agaaccctgg cgcccctgct gccagaaagg
     4861 cctcctccgt gaagcaacaa ccactcgcac gtgtggctcc agtacccggc ctaaatgccg
     4921 cttgattcac cacaaagtgc ttttcagtgc ccccaaactg gattgccaaa cttctgacaa
     4981 gtgtattgtt ccgagtatgg tgagcgaatc ccttctctat tatcaattct cgaaaactct
     5041 tcgggagtca gggcccacgg aaccgagggg gcggatgagg gagaggggga aacaacaatt
     5101 ccgcaccgcg aacaggcacc gctcccctgg acctctccct cagatgtgag gtgcagtcag
     5161 cctttgttgt cattggggtt aagcggaaag gtgtgtgtca gtaggttgtg aggtgaaagc
     5221 gttttcagat gcatagtgag cttaatgtcc ttttcacagt atatcgtgtc tgataggtat
     5281 ctcttattag tatagtcgaa tactagtcaa tagtgcgttt tgtgcaaaat gtccattttg
     5341 tggcagtgat ggggttgttt tatgctattc cgtgtctctg ggtgggcgtg cattgaaaat
     5401 aggggttatc gggtgagtac tgagtttaac atgttctcgt gatcgctgca cgcgccttcg
     5461 agtttttttt ccttttcccc atttttttca acttgaagac ttcaattaca ccaaaaagta
     5521 aaattcacaa gcttgttaac atgattgaac aagatggatt gcacgcaggt tctccggccg
     5581 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5641 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5701 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5761 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5821 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5881 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5941 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6001 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6061 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6121 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6181 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6241 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6301 tcgccttcta tcgccttctt gacgagttct tctgatccgg atcctaagcg gatgcaagcg
     6361 tgtaaagcgc ctcggaggaa cgaaacccct ttgaaaaggt ggctttcgtt tatatattct
     6421 ccatctggtg catcgtgttt atttcgtatt attccctgtc aaaagctgcg gatattcatt
     6481 taatattctt ttcgttatat atttttagtt ctatccttta ttttatccgt tttaatcgtc
     6541 ttctgagacc cacagccctg tagatttctg tgatgtttcg gttgcgtatt ccataatttt
     6601 aagcgtttca cttctatttt ttttcattcc tttgaatttg gatcttaaaa ttatcattgg
     6661 tgccttgtgt tattgtgcgt gctgcgtgtg aatgtggcgc tctgcctgca ggcatgcaag
     6721 ctagcttgta ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
     6781 taattgtagc cgcgttctaa cgacaatatg tacaagccta attgtgtagc atctggctta
     6841 ctgaagcaga ccctatcatc tctctcgtaa actgccgtca gagtcggttt ggttggacga
     6901 accttctgag tttctggtaa cgccgttccg caccccggaa atggtcagcg aaccaatcag
     6961 cagggtcatc gctagccaga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     7021 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     7081 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     7141 gggactgttg ggcgccatct ccttgcacca ttccttgcgg cggcggtgct caacggcctc
     7201 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgatatggt
     7261 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     7321 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     7381 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     7441 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     7501 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7561 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7621 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     7681 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     7741 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7801 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     7861 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     7921 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     7981 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     8041 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     8101 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     8161 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     8221 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     8281 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     8341 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     8401 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     8461 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     8521 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     8581 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8641 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8701 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8761 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     8821 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8881 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8941 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     9001 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     9061 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     9121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     9181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9301 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     9361 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     9421 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     9481 cgattcatta atgcagctgg cttatcgaaa ttaatacgac tcactatagg gagaccg
//
