LOCUS       Exported                4988 bp DNA     circular SYN 24-SEP-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4988)
  AUTHORS   ASTRID HAASE
  TITLE     Direct Submission
  JOURNAL   Exported May 20, 2022 from SnapGene Viewer 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4988
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     protein_bind    505..526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        541..571
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    579..595
                     /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     603..619
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    608..632
                     /label=puc19
     CDS             615..632
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITP"
     misc_feature    632
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    636..1336
                     /label=UP-stream HR
     misc_feature    1116..1135
                     /label=gRNA
     misc_feature    1337..1831
                     /label=NURO UP
     misc_feature    1337..1361
                     /label=ef1ap down
     misc_feature    1362..1368
                     /label=kozak
     primer_bind     1366..1391
                     /label=CMA178/CA20a
     primer_bind     1366..1391
                     /label=CMA178/CA20
     CDS             1369..2163
                     /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     1369..1391
                     /label=CMA203/CA21
     primer_bind     complement(1423..1442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1512..1527
                     /label=AE13
     primer_bind     1593..1607
                     /label=CMA156/CA11
     misc_feature    1806..2298
                     /label=NURO DOWN
     primer_bind     2033..2052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2097..2112
                     /label=AE14
     polyA_signal    2164..2298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2176..2193)
                     /label=TK07
     primer_bind     complement(2201..2220)
                     /label=SV40pA-R
     primer_bind     2228..2248
                     /label=CMA217
     primer_bind     complement(2274..2299)
                     /label=SS204
                     /note="61.8"
     primer_bind     complement(2274..2298)
                     /label=SS037
     primer_bind     complement(2275..2299)
                     /label=CMA266
     primer_bind     complement(2282..2299)
                     /label=SS202
     primer_bind     complement(2282..2299)
                     /label=SS199
     primer_bind     complement(2282..2299)
                     /label=SS197
     primer_bind     complement(2284..2299)
                     /label=CMA101
     misc_feature    2299..2958
                     /label=down-stream HR
     misc_feature    2805..2882
                     /label=EXON3
     CDS             2959..3240
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="LEDPRVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSE
                     EARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     misc_feature    2959..2990
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    2959..2985
                     /label=PUC19
     primer_bind     complement(2991..3007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3481..3585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3586..4446
                     /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"
     rep_origin      join(4617..4988,1..217)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
       61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
      241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
      301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
      361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
      421 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
      481 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      541 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      601 cacaggaaac agctatgacc atgattacgc catgacgggg gcgggtcact cccggacagg
      661 gcccccatga agtttccctg gggactgtgt gactccctag gggtcgggtt acctcccaga
      721 tgggggtggg gtatcacttg cagctggaag gcaggttcat tttaaggtct cgcgggcttc
      781 ggaggcccct ccctcactga gaaaccggct tttgtcgttc ttctgaagcc agagctagtc
      841 tttccaggtg ttagtcgaaa cctcgtggtg cgaccctggt cgtcccaaac cccctaggcc
      901 ttaatcctgg ggcggtgggg gcggggaggc cgtgagcacg gcttccgctc ctccaatccg
      961 ccagagggcg cagcggccgg cctctccctt cccggggttc ttcgcgccgg gccccttccg
     1021 cgtgggtgag tgaatgtgag agtcagcgct cgcgccgcgc gcgccgcccg cctccgctgt
     1081 tcggcgctct gctttaggcg gtggggggcg ggcgcgcgcg taaaagcata gagacaggca
     1141 ttgagctctt gggctagagc gtcgccgagt cggagccgga gcctgagccg cgcgctgtgt
     1201 ctccgctgcg tccgccgagg cccccgagtg tcagggacaa aagcctccgc ctgctcccgc
     1261 agacggggct catctgccgc cgccgccgcg ctgaggagag ttcgccgccg tcgccgcccg
     1321 tgaggatctg agagccaatt agcttgacca agcttgctag caatcaaaat gattgaacaa
     1381 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     1441 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     1501 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     1561 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     1621 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     1681 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1741 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     1801 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     1861 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     1921 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1981 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2041 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2101 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2161 tgaaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2221 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2281 tcttatcatg tctggatcaa gcgccccggg cggaggacct ttcggaagcc gcttagttcg
     2341 caggtgccgc acacttaagt attttgaccc tttcggtgtg ttcttgcagc tggcgaacag
     2401 ctttttcgct aacaaggagt aattcattaa gggtgggggt ggattgggtt ttgaaacgct
     2461 ccaggtcctt agtttcctgt aggtcttaga aggcctttgt ttttcatcct cgtggatcac
     2521 tctctggaag tactctcatg tgatggggga ggggaagcgg gagggaatgg aaatccagag
     2581 gagccaataa aatgaccttt ttagatcaac ttgctctggg tggaggggga cattttcact
     2641 ggcgactgtc gcttcagcat tcactgtcat ctcttaccct tttgaagttc aatcttacgt
     2701 gctttgttaa ctagtaaggt ttttctctgg ttagcatata tgaactaatt tgaaaggaag
     2761 attgtaggat tgggacactc ctaattgaat ttttttttct ttagcaaaat ggatctgtac
     2821 atcaaaagga tggattaaac gatgatgatt ttgaacctta cttgagtcca caggcaaggc
     2881 ccgtgagtag ttaactattt acatatctaa tcgaagggtg tggtatattc tttctccgcc
     2941 ttctaccaat aaatcccact agaggatccc cgggtaccga gctcgaattc actggccgtc
     3001 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     3061 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     3121 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
     3181 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3241 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3301 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3361 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     3421 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     3481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4981 accgaact
//
