LOCUS       pNWA247_(pCMV6-A        6950 bp DNA     circular SYN 19-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 20, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1014..1023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1023..1046
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1065..1472
                     /codon_start=1
                     /label=1-136 RAD23A
                     /translation="MAVTITLKTLQQQTFKIRMEPDETVKVLKEKIEAEKGRDAFPVAG
                     QKLIYAGKILSDDVPIRDYRIDEKNFVVVMVTKTKAGQGTSAPPEASPTAAPESSTSFP
                     PAPTSGMSHPPPAAREDKSPSEESAPTTSPES"
     misc_feature    1389..1905
                     /label=Synthetic fragment
                     /note="cloned into BspEI-MfeI"
     CDS             1473..2084
                     /codon_start=1
                     /label=161-363 RAD23A
                     /translation="SEYETMLTEIMSMGYERERVVAALRASYNNPHRAVEYLLTGIPGS
                     PEPEHGSVQESQVSEQPATEAAGENPLEFLRDQPQFQNMRQVIQQNPALLPALLQQLGQ
                     ENPQLLQQISRHQEQFIQMLNEPPGELADISDVEGEVGAIGEEAPQMNYIQVTPQEKEA
                     IERLKALGFPESLVIQAYFACEKNENLAANFLLSQNFDDE"
     primer_bind     complement(2158..2174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2185..2807
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        2836..3165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3016..3151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3232..4026
                     /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    4200..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4370..4386)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4394..4410
                     /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(4418..4448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4463..4484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4772..5357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5528..6388)
                     /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(6389..6493)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6520..6948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccacca
     1021 tggattacaa ggatgacgac gataaggcga tcgccatgga gcatatggcc gtcactatta
     1081 ccctgaagac cctgcagcag cagacattca agattcggat ggaacctgat gagacagtca
     1141 aagtcctgaa agagaagatc gaggcagaaa aaggacgaga cgcattccct gtcgcaggac
     1201 agaagctgat ctacgctggg aaaattctgt ccgacgatgt gccaatcagg gactatcgca
     1261 ttgatgagaa gaactttgtg gtcgtgatgg tgactaagac caaagccggc caggggacat
     1321 ctgctccccc tgaagcaagt ccaactgccg ctcccgagag ctccacctct ttcccacccg
     1381 ccccaacatc cggaatgtct caccctccac ccgcagcccg cgaagacaaa tccccctctg
     1441 aggaaagcgc ccctaccaca tccccagaga gtagcgagta cgaaaccatg ctgacagaga
     1501 tcatgagtat gggctatgag cgggaaagag tcgtggccgc tctgcgagcc agctacaaca
     1561 atccccaccg ggctgtggag tatctgctga ctgggattcc tggcagcccc gagcctgaac
     1621 atgggtctgt ccaagaaagt caggtgtcag agcagccagc aaccgaggca gcaggagaaa
     1681 acccactgga gttcctgaga gatcagcccc agtttcagaa catgaggcag gtcatccagc
     1741 agaatccagc actgctgcct gctctgctgc agcagctggg ccaggaaaac cctcagctgc
     1801 tgcagcagat ctccaggcat caggagcagt ttattcagat gctgaatgaa cctccagggg
     1861 agctggccga catctctgat gtcgagggcg aagtgggggc aattggagag gaagcccctc
     1921 agatgaatta catccaggtc accccacagg agaaggaagc tattgaacgg ctgaaagcac
     1981 tgggcttccc cgagagcctg gtcatccagg cctattttgc ttgcgagaag aacgaaaatc
     2041 tggctgcaaa cttcctgctg agccagaatt ttgacgatga gtaagaattc ggatccacgc
     2101 gtaagcttct cgagacgcgt taagcggccg cactcgaggt ttaaacggcc ggccgcggtc
     2161 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     2221 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     2281 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     2341 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     2401 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     2461 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     2521 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     2581 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     2641 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     2701 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     2761 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     2821 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3121 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3181 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3241 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3301 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3361 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3421 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3481 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3541 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3601 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3661 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3721 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3781 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3841 tctggattca tcgactgtgg ccggctgggt gtggccgacc gctatcagga catagcgttg
     3901 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3961 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4021 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4081 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4141 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4201 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4261 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4321 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4381 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4441 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4501 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4561 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4621 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4681 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4741 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4801 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4861 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4921 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4981 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5041 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5101 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5161 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5221 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5281 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     6541 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6601 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6661 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6721 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6781 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6841 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6901 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
//
