LOCUS       Exported                6363 bp ds-DNA     circular SYN 16-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 18, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6363
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             913..1851
                     /codon_start=1
                     /label=TEX264, E194A
                     /translation="MSDLLLLGLIGGLTLLLLLTLLAFAGYSGLLAGVEVSAGSPPIRN
                     VTVAYKFHMGLYGETGRLFTESCSISPKLRSIAVYYDNPHMVPPDKCRCAVGSILSEGE
                     ESPSPELIDLYQKFGFKVFSFPAPSHVVTATFPYTTILSIWLATRRVHPALDTYIKERK
                     LCAYPRLEIYQEDQIHFMCPLARQGDFYVPAMKETEWKWRGLVEAIDTQVDGTGADTMS
                     DTSSVSLEVSPGSRETSAATLSPGASSRGWDDGDTRSEHSYSESGASGSSFEELDLEGE
                     GPLGESRLDPGTEPLGTTKWLWEPTAPEKGKE"
     misc_feature    1492..1494
                     /label=E194A
     CDS             1858..1881
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1919..1937)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1963..2187
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2233..2661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2675..3004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2855..2990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3071..3865
                     /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"
     polyA_signal    4039..4160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4209..4225)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4233..4249
                     /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)."
     promoter        complement(4257..4287)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4302..4323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4611..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5367..6227)
                     /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"
     promoter        complement(6228..6332)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccatgtcgga cctgctacta ctgggcctga ttgggggcct gactctctta
      961 ctgctgctga cgctgctggc ctttgccggg tactcagggc tactggctgg ggtggaagtg
     1021 agtgctgggt caccccccat ccgcaacgtc actgtggcct acaagttcca catggggctc
     1081 tatggtgaga ctgggcggct tttcactgag agctgcagca tctctcccaa gctccgctcc
     1141 atcgctgtct actatgacaa cccccacatg gtgccccctg ataagtgccg atgtgccgtg
     1201 ggcagcatcc tgagtgaagg tgaggaatcg ccctcccctg agctcatcga cctctaccag
     1261 aaatttggct tcaaggtgtt ctccttcccg gcacccagcc atgtggtgac agccaccttc
     1321 ccctacacca ccattctgtc catctggctg gctacccgcc gtgtccatcc tgccttggac
     1381 acctacatca aggagcggaa gctgtgtgcc tatcctcggc tggagatcta ccaggaagac
     1441 cagatccatt tcatgtgccc actggcacgg cagggagact tctatgtgcc tgcgatgaag
     1501 gagacagagt ggaaatggcg ggggcttgtg gaggccattg acacccaggt ggatggcaca
     1561 ggagctgaca caatgagtga cacgagttct gtaagcttgg aagtgagccc tggcagccgg
     1621 gagacttcag ctgccacact gtcacctggg gcgagcagcc gtggctggga tgacggtgac
     1681 acccgcagcg agcacagcta cagcgagtca ggtgccagcg gctcctcttt tgaggagctg
     1741 gacttggagg gcgaggggcc cttaggggag tcacggctgg accctgggac tgagcccctg
     1801 gggactacca agtggctctg ggagcccact gcccctgaga agggcaagga gagcgctgac
     1861 tacaaagacg atgacgacaa gtaagcggcc gctcgagcat gcatctagag ggccctattc
     1921 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     1981 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2041 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2101 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2161 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2221 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2281 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2341 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2401 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2461 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2521 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2581 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2641 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2701 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2821 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2881 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2941 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3001 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3061 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3901 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4021 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4081 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4141 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4201 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4261 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4321 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5041 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6361 gtc
//
