LOCUS       Exported                8144 bp ds-DNA     circular SYN 01-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8144)
  AUTHORS   Eric Schreiter
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 5, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8144
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     misc_feature    1774..1830
                     /label=NES
     misc_feature    1774..1777
                     /label=KindOfKozak
     primer_bind     1777..1795
                     /label=f.NES_seq.01.3
     CDS             1837..2370
                     /codon_start=1
                     /label=HaloTag
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEWTLPMAVAR"
     misc_feature    2344..2346
                     /label=G171W
     misc_feature    2359..2361
                     /label=G176A
     misc_feature    2365..2367
                     /label=V178A
     CDS             2371..2430
                     /codon_start=1
                     /label=peptide - 1CDL
                     /translation="ARRKWQKTGHAVRAIGRLSS"
     CDS             2431..2466
                     /codon_start=1
                     /label=Linker
                     /translation="GGSGGSGGSGGS"
     CDS             2467..2907
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(2598..2616)
                     /label=CaM_rev_SeqPrimer
     misc_feature    2727
                     /label=HindIII site removed
     CDS             2908..3261
                     /codon_start=1
                     /label=HaloTag_part
                     /translation="YLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     misc_feature    2908..2910
                     /label=P180Y
     misc_feature    3262..3273
                     /label=linker
     primer_bind     3262..3273
                     /label=Untitled
     primer_bind     3262..3273
                     /label=Untitled
     misc_feature    3274..3993
                     /label=EGFP - codon optimized
     misc_feature    3303
                     /label=NoAgeI_Cut
     intron          4039..4171
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    4180..4768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4651..4662)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4801..4912
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(5008..5026)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5047..5063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_site      5230..5285
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5647..5663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5671..5687
                     /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(5695..5725)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5740..5761
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5820..6015
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5866..6001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6021..6155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6394..6982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7153..8013)
                     /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(8014..8118)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
      901 gtgtgtgggg ggagcggctc ggggggtgcg tgcgtgtgtg tcggcgcggg gctttgtgcg
      961 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     1021 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     1081 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     1141 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     1201 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     1261 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     1321 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     1381 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     1441 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     1501 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg gacggctgcc ttcggggggg
     1561 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
     1621 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
     1681 gtctcatcat tttggcaaag aattcggtac cgcgggcccg gcgcgccagc tggatatctc
     1741 cggacccggg ctagcgcaat tgcggccgcc accatgctgc agaacgagct tgctcttaag
     1801 ttggctggac ttgatattaa caagactgga ggttctgcag aaatcggtac tggctttcca
     1861 ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg
     1921 cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc
     1981 aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg
     2041 ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat
     2101 gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc
     2161 gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg
     2221 gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc
     2281 caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc
     2341 gagtggacgc tgccgatggc cgtcgcgcgc gcccgtcgta aatggcagaa aacaggccat
     2401 gcggttcgtg ctatcggtcg tttgtcgagc ggaggctctg gcggttccgg cggttcaggt
     2461 ggatcagatc aattaacaga ggaacagatt gcggagttta aggaagcgtt ctctttattt
     2521 gataaggatg gcgacggtac aatcactact aaagaattgg gaacagtcat gcgctcattg
     2581 gggcaaaatc cgacagaggc tgaattgcag gacatgatta acgaggtaga cgccgatggg
     2641 aacgggacta tcgactttcc ggaatttctt actatgatgg cacgcaaaat gaaagatacc
     2701 gattctgaag aagagatccg tgaagccttc cgtgtttttg ataaagatgg gaacggctac
     2761 atcagtgctg ctgagttacg ccatgtgatg acaaatctgg gggaaaaact taccgacgaa
     2821 gaagtagacg aaatgattcg cgaggcggat attgacgggg atggacaagt aaactacgag
     2881 gaatttgtgc agatgatgac cgccaagtat ctgactgaag tcgagatgga ccattaccgc
     2941 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
     3001 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
     3061 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
     3121 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
     3181 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg
     3241 tcgacgctcg agatttccgg cggcggaagc ggcatggtct ctaaggggga ggaactcttt
     3301 actggtgtgg tgccaatact tgtagagctc gatggggacg ttaatggaca caaattttct
     3361 gtctcaggtg agggtgaggg agatgcaact tatggaaaac tgacattgaa attcatctgc
     3421 acaacaggca aattgcctgt cccctggcct accctggtaa ccaccctcac ttatggcgtt
     3481 cagtgcttct cccggtatcc tgatcacatg aaacagcatg atttcttcaa atcagcaatg
     3541 cccgaaggtt atgttcaaga gcggaccatc tttttcaagg acgatggaaa ctataaaacc
     3601 agagctgagg ttaagttcga gggagatact ttggtgaatc ggatcgaact gaaggggata
     3661 gacttcaaag aggatgggaa cattttgggg cataaactcg aatataacta caactcccac
     3721 aacgtgtata tcatggctga caaacagaag aacgggatca aagtaaattt taagatacga
     3781 cacaatatag aggacggtag tgtccaactt gctgatcact accagcagaa tacacccatc
     3841 ggagacggac ccgttctctt gcccgataat cactatctct ctacacaatc cgctctgtct
     3901 aaagatccaa acgaaaagcg ggaccacatg gtactgcttg agttcgttac agccgctggt
     3961 atcaccttgg gcatggatga gttgtataaa taaaccggtg ctcgaggtac cgatgaatag
     4021 ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     4081 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     4141 actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa
     4201 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4261 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4321 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4381 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4441 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4501 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4561 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4621 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4681 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4741 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga
     4801 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4861 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4921 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4981 gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     5041 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc
     5101 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     5161 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     5221 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     5281 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     5341 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     5401 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     5461 tgacttgagg ttagattttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     5521 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     5581 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     5641 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5701 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5761 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     5821 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5881 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5941 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6001 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     6061 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6121 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     6181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7141 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7201 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7261 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7321 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7381 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7441 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7501 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7561 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7621 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7681 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7741 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7801 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7861 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7921 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7981 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8041 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8101 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
