LOCUS       Exported                5554 bp ds-DNA     circular SYN 21-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   VB200124-1032nqq
VERSION     .
KEYWORDS    VB200124-1032nqq.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported to SnapGene on Jan 24, 2020
REFERENCE   2  (bases 1 to 5554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 21, 2025 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5554
                     /organism="synthetic DNA construct"
     protein_bind    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        22..1754
                     /label=CAG
                     /note="/application_notes=Strong promoter."
                     /note="/description=CMV early enhancer fused to modified 
                     chicken beta-actin promoter"
                     /note="/full_name=Cytomegalovirus early enhancer fused with
                     chicken beta-actin promoter"
                     /note="/official_designation=CAG"
                     /note="/uuid=420ea94873db443a902370384d17a441"
     enhancer        25..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..683
                     /label=chicken beta-actin promoter
     intron          684..1700
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     protein_bind    1754..1778
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1779..1784
                     /label=Kozak
                     /label=nonstandard type: Miscellaneous
                     /note="/application_notes=Facilitates translation 
                     initiation of ATG start codon downstream of the Kozak 
                     sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
     CDS             1785..3257
                     /codon_start=1
                     /label={hNgn3_5SA}
                     /label=nonstandard type: ORF
                     /note="/application_notes="
                     /note="/official_designation={hNgn3_5SA}"
                     /note="/uuid=864d2defcbe840aeaa0b0a81e6be2e56"
                     /translation="MTPQPSGAPTVQVTRETERSFPRASEDEVTCPTSAPPSPTRTRGN
                     CAEAEEGGCRGAPRKLRARRGGRSRPKSELALSKQRRSRRKKANDRERNRMHNLNSALD
                     ALRGVLPTFPDDAKLTKIETLRFAHNYIWALTQTLRIADHSLYALEPPAPHCGELGAPG
                     GAPGDWGSLYAPVSQAGSLAPAASLEERPGLLGATFSACLAPGSLAFSDFLGSAGSAAG
                     SGEFDYKDHDGDYKDHDIDYKDDDDKGTNTPGINLIKEDMRVKVHMEGNVNGHAFVIEG
                     EGKGKPYEGTQTANLTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEG
                     YSWERTMTFEDKGICTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEK
                     LHVRDGLLVGNINMALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDY
                     NKVKLYEHAVARYSPLPSQVWGTYPYDVPDYA"
     CDS             1785..2426
                     /codon_start=1
                     /label=5SA-Ngn3
                     /translation="MTPQPSGAPTVQVTRETERSFPRASEDEVTCPTSAPPSPTRTRGN
                     CAEAEEGGCRGAPRKLRARRGGRSRPKSELALSKQRRSRRKKANDRERNRMHNLNSALD
                     ALRGVLPTFPDDAKLTKIETLRFAHNYIWALTQTLRIADHSLYALEPPAPHCGELGAPG
                     GAPGDWGSLYAPVSQAGSLAPAASLEERPGLLGATFSACLAPGSLAFSDFL"
     CDS             2427..2462
                     /codon_start=1
                     /product="From WALDO ET AL 
                     http://www.nature.com/nbt/journal/v17/n7/full/nbt0799_691.h
                     tml"
                     /label=Linker
                     /translation="GSAGSAAGSGEF"
     CDS             2463..2528
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2535..3221
                     /codon_start=1
                     /product="monomeric photoswitchable green-to-red 
                     fluorescent protein from Dendronephthya"
                     /label=Dendra2
                     /note="mammalian codon-optimized"
                     /translation="NTPGINLIKEDMRVKVHMEGNVNGHAFVIEGEGKGKPYEGTQTAN
                     LTVKEGAPLPFSYDILTTAVHYGNRVFTKYPEDIPDYFKQSFPEGYSWERTMTFEDKGI
                     CTIRSDISLEGDCFFQNVRFKGTNFPPNGPVMQKKTLKWEPSTEKLHVRDGLLVGNINM
                     ALLLEGGGHYLCDFKTTYKAKKVVQLPDAHFVDHRIEILGNDSDYNKVKLYEHAVARYS
                     PLPSQVW"
     CDS             3228..3254
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3228..3254
                     /label=HA tag
                     /label=insertion_seq
                     /note="vntifkey=14"
     polyA_signal    3302..3523
                     /label=SV40 late pA
                     /note="/application_notes=Allows transcription termination 
                     and polyadenylation of mRNA transcribed by Pol II RNA 
                     polymerase."
                     /note="/description=Simian virus 40 late polyadenylation 
                     signal"
                     /note="/official_designation=SV40 late pA"
                     /note="/uuid=d23bf533e0924dfcb932fc3ddd77e791"
     polyA_signal    3311..3432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3523..3524
                     /label=No marker
                     /label=nonstandard type: Marker
                     /note="/application_notes="
                     /note="/official_designation=No marker(ns)"
                     /note="/uuid=72e1edd2c1a540b28efbb477521f8696"
     rep_origin      complement(3719..4307)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E. coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=41c627938a364ebfa136b55444385c5b"
     misc_feature    complement(4478..5338)
                     /label=AmpiciIIin
                     /label=nonstandard type: ORF
                     /note="/application_notes=Allows E. coli to be resistant to
                     ampiciIIin."
                     /note="/description=AmpiciIIin resistance gene"
                     /note="/full_name=Ampicillin resistance gene"
                     /note="/official_designation=Ampicillin"
                     /note="/uuid=0bc7d58885794858b29e98fe171ff1d5"
     CDS             complement(4478..5338)
                     /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(5339..5443)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caactttgta tagaaaagtt gctcgacatt gattattgac tagttattaa tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      421 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      481 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
      541 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      601 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      661 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      721 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      781 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      841 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      901 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
      961 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     1021 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     1081 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     1141 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     1201 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     1261 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     1321 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     1381 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     1441 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     1501 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     1561 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     1621 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     1681 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1741 ttggcaaaga attgcaagtt tgtacaaaaa agcaggctgc caccatgacg cctcaaccct
     1801 cgggtgcgcc cactgtccaa gtgacccgtg agacggagcg gtccttcccc agagcctcgg
     1861 aagacgaagt gacctgcccc acgtccgccc cgcccagccc cactcgcaca cgggggaact
     1921 gcgcagaggc ggaagaggga ggctgccgag gggccccgag gaagctccgg gcacggcgcg
     1981 ggggacgcag ccggcctaag agcgagttgg cactgagcaa gcagcgacgg agtcggcgaa
     2041 agaaggccaa cgaccgcgag cgcaatcgaa tgcacaacct caactcggca ctggacgccc
     2101 tgcgcggtgt cctgcccacc ttcccagacg acgcgaagct caccaagatc gagacgctgc
     2161 gcttcgccca caactacatc tgggcgctga ctcaaacgct gcgcatagcg gaccacagct
     2221 tgtacgcgct ggagccgccg gcgccgcact gcggggagct gggcgcccca ggcggtgccc
     2281 ccggggactg ggggtccctc tacgccccag tctcccaggc tggcagcctg gctcccgccg
     2341 cgtcgctgga ggagaggccc gggctgctgg gggccacctt ttccgcctgc ttggccccag
     2401 gcagtctggc tttctcagat tttctgggct ccgctggctc cgctgctggt tctggcgaat
     2461 ttgactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     2521 atgacaaggg taccaacacc ccgggaatta acctgatcaa ggaggacatg cgcgtgaagg
     2581 tgcacatgga gggcaacgtg aacggccacg ccttcgtgat cgagggcgag ggcaagggca
     2641 agccctacga gggcacccag accgccaacc tgaccgtgaa ggagggcgcc cccctgccct
     2701 tcagctacga catcctgacc accgccgtgc actacggcaa ccgggtgttc accaagtacc
     2761 ccgaggacat ccccgactac ttcaagcaga gcttccccga gggctacagc tgggagcgca
     2821 ccatgacctt cgaggacaag ggcatctgca ccatccgcag cgacatcagc ctggagggcg
     2881 actgcttctt ccagaacgtg cgcttcaagg gcaccaactt cccccccaac ggccccgtga
     2941 tgcagaagaa gaccctgaag tgggagccca gcaccgagaa gctgcacgtg cgcgacggcc
     3001 tgctggtggg caacatcaac atggccctgc tgctggaggg cggcggccac tacctgtgcg
     3061 acttcaagac cacctacaag gccaagaagg tggtgcagct gcccgacgcc cacttcgtgg
     3121 accaccgcat cgagatcctg ggcaacgaca gcgactacaa caaggtgaag ctgtacgagc
     3181 acgccgtggc ccgctacagc cccctgccca gccaggtgtg gggtacctac ccatacgatg
     3241 ttccagatta cgcttgaacc cagctttctt gtacaaagtg gtgatggccg gccgcttcga
     3301 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3361 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3421 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     3481 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtagcggccg cggcgctctt
     3541 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3601 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3661 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctctct tcgggaagcg
     3901 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4081 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4141 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4201 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4261 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4321 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4381 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4441 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4501 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4561 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4621 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4681 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4741 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4801 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4861 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4921 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4981 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5041 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5101 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5161 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5221 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5281 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5341 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5401 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5461 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     5521 tcacgaggcc ctttcgtcgg cgcgccgcgg ccgc
//
