LOCUS       DC391-pBO101_-_p        6401 bp DNA     circular SYN 22-APR-2023
DEFINITION  Ligation of 2 sequences.
ACCESSION   urn.local...as2-e7pntkb
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 14, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          393..893
                     /note="organism: synthetic DNA construct mol_type: other
                     DNA"
     source          join(3375..6401,894..941,942..3113,1..389)
                     /mol_type="other DNA"
                     /label=/NCBI Join Type=join
                     /note="/NCBI Join Type=join"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /standard_name="CMV promoter"
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     287..307
                     /standard_name="CMV-F"
                     /label=CMV-F
     primer_bind     333..357
                     /standard_name="LNCX"
                     /label=LNCX
     primer_bind     complement(347..368)
                     /standard_name="oBO68"
                     /label=oBO68
     primer_bind     347..368
                     /standard_name="oBO69"
                     /label=oBO69
     primer_bind     complement(348..367)
                     /standard_name="Express R"
                     /label=Express R
     CDS             390..392
                     /codon_start=1
                     /standard_name="AID*∆"
                     /label=AID*∆
                     /translation="M"
     misc_feature    393..893
                     /standard_name="GACAGCCTCTTGATGAACCGGAGGGAGTTTCTTTACCAATTC 
                     AAAAATGTCCGCTGGGCTAAGGGTCGGCGTGAGACCTACCTGTGCTACGTAGTGAAGA 
                     GGCGTGACAGTGCTACATCCTTTTCACTGGACTTTGGTTATCTTCGCAATAAGAACGG 
                     CTGCCACGTGGAATTGCTCTTCCTCCGCTACATCTCGGACTGGGACCTAGACCCTGGC 
                     CGCTGCTACCGCGTCACCTGGTTCATCTCCTGGAGCCCCTGCTACGACTGTGCCCGAC 
                     ATGTGGCCGACTTTCTGCGAGGGAACCCCAACCTCAGTCTGAGGATCTTCACCGCGCG 
                     CCTCTACTTCTGTGAGGACCGCAAGGCTGAGCCCGAGGGGCTGCGGCGGCTGCACCGC 
                     GCCGGGGTGCAAATAGCCATCATGACCTTCAAAGATTATTTTTACTGCTGGAATACTT 
                     TTGTAGAAAACCACGGAAGAACTTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCAGT 
                     TCGTCTCTCCAGACAGCTTCGGCGCATCCTTTTGCCCCTGTATGAGGTTGATGACTTA 
                     CGAGACGCATTTCGTACT"
                     /label=nonstandard type: Editing History Replacement
     CDS             393..893
                     /standard_name="Translation 90-686"
                     /label=Translation 90-686
     misc_feature    complement(393..395)
                     /standard_name="68"
                     /label=68
     misc_feature    393..395
                     /standard_name="overlap"
                     /label=overlap
     misc_feature    393..395
                     /standard_name="61"
                     /label=61
     misc_feature    396..893
                     /standard_name="TadDE"
                     /label=TadDE
     CDS             894..941
                     /codon_start=1
                     /standard_name="XTEN"
                     /label=XTEN
                     /translation="SGSETPGTSESATPES"
     primer_bind     894..910
                     /standard_name="oBO67"
                     /label=oBO67
     primer_bind     894..908
                     /standard_name="oBO172"
                     /label=oBO172
     primer_bind     894..908
                     /standard_name="oBO176"
                     /label=oBO176
     primer_bind     complement(902..918)
                     /standard_name="oBO71"
                     /label=oBO71
     primer_bind     complement(924..941)
                     /standard_name="oBO114"
                     /label=oBO114
     primer_bind     complement(925..961)
                     /standard_name="oBO177 - GsPcrA-M6/XTEN_R"
                     /label=oBO177 - GsPcrA-M6/XTEN_R
     primer_bind     complement(925..961)
                     /standard_name="oBO177"
                     /label=oBO177
     primer_bind     complement(925..941)
                     /standard_name="XTEN_R"
                     /label=XTEN_R
     primer_bind     complement(926..941)
                     /standard_name="oBO173"
                     /label=oBO173
     CDS             942..3113
                     /codon_start=1
                     /standard_name="GsPcrA-M6"
                     /label=GsPcrA-M6
                     /translation="MNFLSEQLLAHLNKEQQEAVRTTEGPLLIMAGAGSGKTRVLTHRI
                     AYLMAEKHVAPWNILAITFTNKAAREMRERVQSLLGGAAEDVWISTFASMAVRILRRDI
                     DRIGINRNFSILDPTDQLSVMKTILKEKNIDPKKFEPRTILGTISAAKNELLPPEQFAK
                     RASTYYEKVVSDVYQEYQQRLLRCHSLDFDDLIMTTIQLFDRVPDVLHYYQYKFQYIHI
                     DEYQDTNRAQYTLVKKLAERFQNIAAVGDADQSIYRWRGADIQNILSFERDYPNAKVIL
                     LEQNYRSTKRILQAANEVIEHNVNRKPKRIWTENPEGKPILYYEAMNEADEAQFVAGRI
                     REAVERGERRYRDFAVLYRTNAQSRVMEEMLLKANIPYQIVGGVKFYDRKEIKDILAYL
                     RVIANPDDDCSLLRIINVPKRGIGASTIDKLVRYAADHELSLFEALGELEMIGLGAKAA
                     GALAAFRSQLEQWTQLQEYVSVTELVEEVLDKSGYREMLKAERTIEAQSRLENLDEFLS
                     VTKHFENVSDDKSLIAFLTDLALISDLDELDGTEQAAEGDAVMLMTLHAAKGLEFPVVF
                     LIGMEEGIFPHNRSLEDDDEMEEERRLAYVGITRAEEELVLTSAQMRTLFGNIQMDPPS
                     RFLNEIPAHLLETASRRQAGASRPAVSRPQASGAVGSWKVGDRANHRKWGIGTVVSVRG
                     GGDDQELDIAFPSPIGIKRLLAKFAPIEKV"
     misc_feature    1218..1220
                     /standard_name="H93A"
                     /label=H93A
     misc_feature    1227..1229
                     /standard_name="C96A"
                     /label=C96A
     misc_feature    1500..1502
                     /standard_name="N187C"
                     /label=nonstandard type: misc
     misc_feature    1680..1682
                     /standard_name="C247A"
                     /label=nonstandard type: misc
     misc_feature    2091..2093
                     /standard_name="L834V"
                     /label=nonstandard type: misc
     CDS             2166..2168
                     /codon_start=1
                     /standard_name="L409C"
                     /label=L409C
                     /translation="C"
     primer_bind     complement(3105..3125)
                     /standard_name="GsPcrA-M6-SGGS_R"
                     /label=GsPcrA-M6-SGGS_R
     CDS             3126..3374
                     /standard_name="UGI"
                     /label=UGI
     primer_bind     3126..3160
                     /standard_name="UGI_F"
                     /label=UGI_F
     primer_bind     complement(3349..3385)
                     /standard_name="oBO57"
                     /label=oBO57
     primer_bind     complement(3350..3374)
                     /standard_name="oBO171"
                     /label=oBO171
     primer_bind     3373..3398
                     /standard_name="oBO169"
                     /label=oBO169
     primer_bind     3375..3395
                     /standard_name="no_ugi"
                     /label=no_ugi
     primer_bind     3375..3395
                     /standard_name="oBO112"
                     /label=oBO112
     primer_bind     3387..3413
                     /standard_name="bp/NLS_F"
                     /label=bp/NLS_F
     primer_bind     3387..3413
                     /standard_name="SV40_NLS_F"
                     /label=SV40_NLS_F
     CDS             3387..3407
                     /codon_start=1
                     /standard_name="SV40 NLS"
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     3408..3431
                     /standard_name="oBO72"
                     /label=oBO72
     CDS             3416..3433
                     /codon_start=1
                     /standard_name="6xHis"
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3435..3456
                     /standard_name="5,565 F"
                     /label=5,565 F
     primer_bind     3435..3456
                     /standard_name="Express Fwd"
                     /label=Express Fwd
     primer_bind     complement(3457..3474)
                     /standard_name="BGH-rev"
                     /label=BGH-rev
     polyA_signal    3462..3686
                     /label=bovine growth hormone polyadenylation signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="/standard_name=bGH poly(A) signal"
                     /note="/standard_name=bGH poly(A) signal"
     primer_bind     complement(3462..3482)
                     /standard_name="oBO32"
                     /label=oBO32
     primer_bind     complement(3529..3548)
                     /standard_name="5,678 R"
                     /label=5,678 R
     primer_bind     complement(3719..3733)
                     /standard_name="pBluescriptKS"
                     /label=pBluescriptKS
     primer_bind     complement(3758..3774)
                     /standard_name="M13 rev"
                     /label=M13 rev
     primer_bind     complement(3758..3774)
                     /standard_name="M13 Reverse"
                     /label=M13 Reverse
     primer_bind     complement(3771..3793)
                     /standard_name="M13/pUC Reverse"
                     /label=M13/pUC Reverse
     protein_bind    3781..3797
                     /standard_name="lac operator"
                     /label=lac operator binding site
                     /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(3805..3835)
                     /standard_name="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3871
                     /standard_name="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
     primer_bind     complement(3989..4006)
                     /standard_name="L4440"
                     /label=L4440
     rep_origin      complement(4159..4747)
                     /direction=LEFT
     primer_bind     complement(4240..4259)
                     /standard_name="pBR322ori-F"
                     /label=pBR322ori-F
     CDS             complement(4918..5778)
                     /codon_start=1
                     /gene="bla"
                     /standard_name="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5542..5561
                     /standard_name="Amp-R"
                     /label=Amp-R
     promoter        complement(5779..5883)
                     /gene="bla"
                     /standard_name="AmpR promoter"
                     /label=AmpR promoter
     primer_bind     complement(5963..5982)
                     /standard_name="pRS-marker"
                     /label=pRS-marker
     enhancer        join(6153..6401,1..131)
                     /standard_name="CMV enhancer"
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgcgaga gccgccacca tgagttctga ggtggagttt tcccacgagt
      421 actggatgag acatgccctg accctggcca agagggcacg ggatgaggga gaggcgcctg
      481 tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac agacgtatcg
      541 gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc ggcctggtca
      601 tgcagaactc cagactgatt gacgccaccc tgtacgtgac attcgagcct tgcgtgatgt
      661 gcgccggcgc catgatcaac tctaggatcg gccgcgtggt gtttggcgtg aggaactcaa
      721 aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg aatcaccgcg
      781 tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc gatttctatc
      841 ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc aacagcggca
      901 gcgagactcc cgggacctca gagtccgcca cacccgaaag tatgaacttc ctgtctgagc
      961 aactgctggc acacctgaac aaggagcagc aagaagctgt gcggaccacc gagggacctc
     1021 tgctgatcat ggccggcgct ggaagcggaa aaacaagagt gctgacacac cgcatcgcct
     1081 acctgatggc tgagaagcac gtggcccctt ggaacatcct ggccattacc tttacaaaca
     1141 aggccgctag agagatgaga gaaagagtgc agagcctcct gggaggcgcc gccgaggacg
     1201 tgtggatcag caccttcgcc agcatggccg tgcggatcct gagaagagat atcgacagaa
     1261 tcggcatcaa ccggaacttc agcatcctgg acccaacaga ccagctgagc gtgatgaaaa
     1321 ccatcctgaa agaaaagaac atcgacccca agaaattcga gcctagaaca atcctgggca
     1381 caatcagcgc cgccaagaat gaactgctgc ctccagaaca gtttgcgaag cgggcctcca
     1441 cctactatga gaaagtggtg tctgacgtct accaggagta tcagcagcgg ctgctcaggt
     1501 gtcacagcct tgacttcgat gatctgatca tgaccacaat ccagctgttt gaccgggtcc
     1561 ccgacgtgct ccactactac caatacaagt ttcagtacat ccacatcgat gagtaccagg
     1621 acacaaacag agcccaatac accctggtga aaaagctggc tgagcggttc cagaacatcg
     1681 ccgccgtggg cgacgccgat cagtctatct acagatggcg gggcgccgac atccagaaca
     1741 tcctgagctt cgaaagagat taccccaacg ccaaagtgat cctgctggag caaaattacc
     1801 ggagcacgaa gcgcatcctg caggccgcaa acgaggtgat cgagcacaac gtgaacagaa
     1861 agcctaagcg gatctggacc gagaatcctg agggcaagcc catcctgtac tacgaggcca
     1921 tgaacgaagc cgacgaggcc cagttcgtgg ccggcagaat cagagaggcc gtggagcgcg
     1981 gcgagagaag ataccgagac ttcgccgtgc tgtacagaac caatgcccag tccagagtca
     2041 tggaagagat gctgctgaag gccaacatcc cttaccagat agtgggcggc gtgaagttct
     2101 acgacagaaa ggaaatcaag gacatcctgg cttatctgcg ggtcatcgca aacccagacg
     2161 acgactgcag cctgctgaga attatcaacg ttcctaaaag aggcatcgga gcctctacca
     2221 tcgacaagct ggtccggtac gccgccgacc acgagctgag cctgttcgag gccctgggcg
     2281 agctggagat gatcggtctt ggcgccaagg ccgctggggc cctggccgcc ttcagaagcc
     2341 agctggaaca gtggacacag ctgcaggagt acgtgagcgt gaccgaactg gtggaagagg
     2401 tgctggacaa gtctggctac cgggaaatgc tgaaggccga aagaaccatc gaagcccaga
     2461 gcagactgga gaacctggat gagttcctga gcgtgaccaa gcatttcgaa aacgtgtccg
     2521 acgacaagag cctgatcgcc ttcctgaccg atctggctct gatcagcgac ctggacgagc
     2581 tggatggcac cgagcaggcc gccgagggcg acgccgtgat gctgatgacc ctgcacgccg
     2641 ccaagggcct ggaattcccc gtggtctttc tgataggcat ggaagaaggc atcttccctc
     2701 acaatagaag cctggaggat gacgatgaga tggaagagga gagaagactg gcctacgtgg
     2761 gcatcaccag agctgaagag gaactggtgc tgaccagcgc ccagatgcgg accctgttcg
     2821 gcaacattca gatggaccct cctagccggt tccttaacga gatccccgcc catctgcttg
     2881 agacagccag cagacggcag gccggagctt ccaggcctgc cgtgagcaga ccccaggcct
     2941 ctggcgctgt gggcagctgg aaggtgggcg atcgggcaaa tcacagaaag tggggcatcg
     3001 gaaccgtggt ttccgtgcgg ggcggcggag atgatcagga actggacatc gcttttccat
     3061 ctcctatcgg cattaagaga ctgctagcta aattcgcccc tatcgagaag gtgtctggtg
     3121 gttctactaa tctgtcagat attattgaaa aggagaccgg taagcaactg gttatccagg
     3181 aatccatcct catgctccca gaggaggtgg aagaagtcat tgggaacaag ccggaaagcg
     3241 atatactcgt gcacaccgcc tacgacgaga gcaccgacga gaatgtcatg cttctgacta
     3301 gcgacgcccc tgaatacaag ccttgggctc tggtcataca ggatagcaac ggtgagaaca
     3361 agattaagat gctctctggt ggttctccca agaagaagag gaaagtctaa ccggtcatca
     3421 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3481 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3541 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3601 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3661 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     3721 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3781 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     3841 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3901 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3961 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4021 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4981 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5041 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5101 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5161 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5221 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5281 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5341 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5401 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5461 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5521 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5581 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5641 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5701 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5761 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5821 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5881 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
     5941 tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6001 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6061 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     6181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6361 tttacggtaa actgcccact tggcagtaca tcaagtgtat c
//
