LOCUS       pNWA272_(pCMV6-A        6923 bp DNA     circular SYN 05-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6923)
  AUTHORS   nichd_dir_cssc_sub
  TITLE     Direct Submission
  JOURNAL   Exported Apr 24, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6923
                     /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"
     misc_feature    1047..1220
                     /label=Synthetic fragment
                     /note="Cloned into AsiSI-AfeI of pNWA41"
     CDS             1065..2054
                     /codon_start=1
                     /label=80-409 RAD23B CDS
                     /translation="VSTPAPATTQQSAPASTTAVTSSTTTTVAQAPTPVPALAPTSTPA
                     SITPASATASSEPAPASAAKQEKPAEKPAETPVATSPTATDSTSGDSSRSNLFEDATSA
                     LVTGQSYENMVTEIMSMGYEREQVIAALRASFNNPDRAVEYLLMGIPGDRESQAVVDPP
                     QAASTGVPQSSAVAAAAATTTATTTTTSSGGHPLEFLRNQPQFQQMRQIIQQNPSLLPA
                     LLQQIGRENPQLLQQISQHQEHFIQMLNEPVQEAGGQGGGGGGGSGGIAEAGSGHMNYI
                     QVTPQEKEAIERLKALGFPEGLVIQAYFACEKNENLAANFLLQQNFDED"
     misc_feature    1389..1511
                     /label=UBA1
     misc_feature    1653..1844
                     /label=XPC-binding
     misc_feature    1917..2039
                     /label=UBA2
     primer_bind     complement(2131..2147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2158..2780
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        2809..3138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2989..3124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3205..3999
                     /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    4173..4294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4343..4359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4367..4383
                     /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(4391..4421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4436..4457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4745..5330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5501..6361)
                     /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(6362..6466)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6493..6921
                     /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 gcatgtgtcc accccagctc
     1081 ccgcaaccac acagcagtct gccccagcta gtactaccgc cgtgacaagc tccacaacta
     1141 ccacagtcgc acaggcacct actccagtgc cagctctggc acccacatca actcctgcca
     1201 gcatcacacc agccagcgct actgcatcta gtgaacctgc cccagcttcc gccgctaagc
     1261 aggagaaacc agctgaaaaa cccgcagaga ctcctgtggc cacctcaccc accgctacag
     1321 acagcacctc cggcgattca agccggtcca acctgttcga ggacgctacc tccgcactgg
     1381 tcacaggcca gtcttacgaa aatatggtga cagagatcat gtctatgggg tatgagcggg
     1441 aacaggtcat tgcagccctg agagctagtt ttaacaatcc cgatagggca gtggaatacc
     1501 tgctgatggg catccctggg gacagagagt ctcaggcagt cgtggacccc cctcaggctg
     1561 caagtaccgg agtcccacag tcctctgcag tggcagctgc agcagctact accacagcta
     1621 ctaccacaac taccagttca ggcgggcacc ctctggagtt cctgcgcaac cagccacagt
     1681 ttcagcagat gcgacagatc attcagcaga atccatccct gctgccagca ctgctgcagc
     1741 agattggacg cgaaaacccc cagctgctgc agcagatctc tcagcaccag gagcatttca
     1801 ttcagatgct gaatgaacct gtgcaggagg caggaggaca gggaggagga ggaggaggcg
     1861 ggagtggagg catcgcagag gccgggtcag gacacatgaa ctacatccag gtcacaccac
     1921 aggagaagga agcaattgaa aggctgaaag ccctgggatt ccccgagggc ctggtcatcc
     1981 aggcctattt tgcttgcgag aagaacgaaa atctggcagc caacttcctg ctgcagcaga
     2041 attttgacga ggattaagaa ttcggatcca cgcgtaagct tctcgagacg cgttaagcgg
     2101 ccgcactcga ggtttaaacg gccggccgcg gtcatagctg tttcctgaac agatcccggg
     2161 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     2221 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     2281 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     2341 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     2401 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg
     2461 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     2521 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     2581 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt
     2641 ttaaaataac tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc
     2701 caaccggtgg gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact
     2761 cagtagatgc ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta
     2821 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2881 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2941 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3001 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3061 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3121 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3181 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggccg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4021 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4081 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4141 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4201 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4261 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4321 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4381 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4441 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4501 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4561 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4621 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4681 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4741 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4801 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4861 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4921 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4981 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5041 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5101 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5161 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5221 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5281 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5341 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5401 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5461 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5521 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5581 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5641 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5701 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5761 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5821 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5881 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5941 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6001 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6061 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6121 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6181 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6241 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6301 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6361 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6421 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6481 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6541 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6601 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6661 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6721 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6781 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6841 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6901 aacaaaaatt taacgcgaat ttt
//
