LOCUS       Exported                5974 bp ds-DNA     circular SYN 03-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5974)
  AUTHORS   Baylor-NRI
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5974
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     170..189
                     /label=oJDW 855
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     exon            662..751
                     /label=chicken B-actin exon
     intron          752..1670
                     /label=chicken b-actin intron + rabbit b-globin intron
     misc_feature    1594..1616
                     /label=pCAGGS FWDSeq
     primer_bind     1594..1616
                     /label=oJDW 972
     intron          1671..1723
                     /label=rabbit B-globin intron
     protein_bind    1737..1761
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1772..1777
                     /label=TOPO Site 5'
     misc_feature    1782..1790
                     /label=Kozak
     CDS             1791..2834
                     /codon_start=1
                     /label=DRE recombinase
                     /translation="MGASELIISGSSGGFLRNIGKEYQEAAENFMRFMNDQGAYAPNTL
                     RDLRLVFHSWARWCHARQLAWFPISPEMAREYFLQLHDADLASTTIDKHYAMLNMLLSH
                     CGLPPLSDDKSVSLAMRRIRREAATEKGERTGQAIPLRWDDLKLLDVLLSRSERLVDLR
                     NRAFLFVAYNTLMRMSEISRIRVGDLDQTGDTVTLHISHTKTITTAAGLDKVLSRRTTA
                     VLNDWLDVSGLREHPDAVLFPPIHRSNKARITTTPLTAPAMEKIFSDAWVLLNKRDATP
                     NKGRYRTWTGHSARVGAAIDMAEKQVSMVEIMQEGTWKKPETLMRYLRRGGVSVGANSR
                     LMDSASGA"
     misc_feature    2835..2861
                     /label=NLS-stop
     CDS             2838..2858
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2865..2870
                     /label=TOPO SITE 3'
     protein_bind    complement(2881..2905)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2995..3012)
                     /label=oJDW 748
     polyA_signal    3062..3117
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3476..3492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3500..3516
                     /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(3524..3554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3569..3590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3648..3844
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3695..3830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3850..3984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(3887..3906)
                     /label=SV40pA REVSeq
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4982..5842)
                     /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(5843..5947)
                     /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 gcgaggggcg
      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 cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcctcgag gatatcacaa
     1741 gtttgtacaa aaaagcaggc tccgcggccg cccccttcac cgtcgccacc atgggtgcta
     1801 gcgagctgat catctctggc tcctctggag gattcctgag gaacatcggc aaggagtacc
     1861 aggaggctgc tgagaacttc atgagattca tgaatgacca gggagcctac gcccctaaca
     1921 ccctgagaga cctgaggctg gtgttccact cctgggctag atggtgccac gctagacagc
     1981 tggcctggtt ccctatctct cctgagatgg ctagggagta cttccttcag ctgcacgatg
     2041 ctgacctggc ctctaccacc atcgacaagc actacgccat gctgaacatg ctgctgtccc
     2101 actgtggcct gcctcctctg tctgatgaca agtctgtgag cctggccatg aggagaatcc
     2161 ggagagaggc tgccaccgag aagggagaga gaaccggcca ggccatccct ctgagatggg
     2221 atgacctgaa gctgctggat gtgctgctgt ctagatctga gagactggtg gacctgagga
     2281 atagggcctt cctgtttgtg gcctacaaca ccctgatgag gatgtctgag atctctagga
     2341 tcagagtggg agacctggac cagaccggag acaccgtgac cctgcacatc tcccacacca
     2401 agaccatcac caccgctgct ggcctggaca aagtgctgtc taggaggacc accgctgtgc
     2461 tgaatgactg gctggatgtg tctggcctga gagagcaccc tgacgctgtg ctgttccctc
     2521 ctatccaccg gagcaacaag gctaggatca ccaccacccc tctgaccgcc cctgccatgg
     2581 agaagatttt tagcgatgcc tgggtgctgc tgaacaagag ggatgccacc cctaacaagg
     2641 gccgctaccg gacctggacc ggccactctg ctagagtggg agctgccatc gacatggctg
     2701 agaagcaagt gtccatggtg gagatcatgc aggagggcac ctggaaaaag cctgagacac
     2761 tgatgagata cctgaggagg ggaggagtgt ctgtgggagc caactctagg ctgatggact
     2821 ccgctagcgg cgccggtcct aagaagaaga ggaaagtgtg atgaaagggt gggcgcgccg
     2881 acccagcttt cttgtacaaa gtggtgatat cgcggccgca ctcctcaggt gcaggctgcc
     2941 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     3001 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     3061 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3121 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     3181 ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca
     3241 gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg
     3301 ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc
     3361 cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt
     3421 ccctcttctc ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat
     3481 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3541 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3601 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     3661 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3721 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3781 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3841 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3961 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     4021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5941 ttccgcgcac atttccccga aaagtgccac ctga
//
