LOCUS       Exported                7088 bp DNA     circular SYN 25-AUG-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 2, 2023 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7088
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(941..1801)
                     /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(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2567..2731
                     /label=ITR
     polyA_signal    2855..2976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3006..3022
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3025..3613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3131..3142
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3710..3757
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3784..3824
                     /label=Fragment.FOR (1)
     regulatory      3793..3802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3799..3861
                     /label=Ig K-Chain
                     /note="Ig K-Chain"
     CDS             3799..3861
                     /codon_start=1
                     /label=Feature 29
                     /translation="METDTLLLWVLLLWVPGSTGD"
     sig_peptide     3799..3861
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             3838..3861
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="WVPGSTGD"
     primer_bind     complement(3841..3861)
                     /label=Vector.REV
     CDS             3862..5424
                     /codon_start=1
                     /translation="GSAAGSTLDKIAKNGVIVVGHRDSSVPFSYYDSQHKVVGYSQEYS
                     NAIVEAVKKKLNKPDLQVKLIPITSQNRIPLLQNGTYDFECGSTTNNVERQKQAAFSDT
                     IFVVGTRLLTKKGGDIKDFANLKDKAVVVTSGTTSEVLLNKLNEEQKMNMRIISAKDHG
                     DSFRTLESGRAVAFMMDDSLLAGERAKAKKPDDWEIVGKPQSQEAYGCMLRKDDPQFKK
                     LMDDTIAQVQTSGEAEKWFDKWFKNPILVNHTVYITADKQKNGIKANFKIHHNVEDGSV
                     QLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLEFVTATGNSLGMDE
                     LYKGGTGGSMSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTT
                     GKLPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTR
                     AEVKLEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNNPLNMNFELSDEMKALFKEPND
                     KALK"
     CDS             3862..4614
                     /codon_start=1
                     /label=ybeJ1-253
                     /note="ybeJ1-253"
                     /translation="GSAAGSTLDKIAKNGVIVVGHRDSSVPFSYYDSQHKVVGYSQEYS
                     NAIVEAVKKKLNKPDLQVKLIPITSQNRIPLLQNGTYDFECGSTTNNVERQKQAAFSDT
                     IFVVGTRLLTKKGGDIKDFANLKDKAVVVTSGTTSEVLLNKLNEEQKMNMRIISAKDHG
                     DSFRTLESGRAVAFMMDDSLLAGERAKAKKPDDWEIVGKPQSQEAYGCMLRKDDPQFKK
                     LMDDTIAQVQTSGEAEKWFDKWFKNPILV"
     primer_bind     complement(4845..4865)
                     /label=Primer 4a
     misc_feature    4891..4908
                     /label=cpsfGFP
                     /note="cpsfGFP"
     misc_feature    4909..5346
                     /label=cpSF Venus
                     /note="cpSF Venus"
     misc_feature    5347..5424
                     /label=YbeJ254-280
                     /note="YbeJ254-280"
     primer_bind     5425..5447
                     /label=Vector.FOR
     primer_bind     complement(5436..5466)
                     /label=299-R-OpenVec23in4
     CDS             5437..5466
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(5441..5466)
                     /label=Primer 1
     CDS             5467..5819
                     /codon_start=1
                     /label=gBlock replacing part
                     /translation="NAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLW
                     QKKPRDRHKQLRATARATDYLQADAITRIPDYRYRYQRRDRDDDRDTEPDHDRDADPVG
                     VKGFNTLPDTEID"
     CDS             5467..5529
                     /codon_start=1
                     /label=PDGFR-beta hinge
                     /translation="NAVGQDTQEVIVVPHSLPFKV"
     CDS             5530..5592
                     /codon_start=1
                     /label=PDGFR-beta TM domain
                     /translation="VVISAILALVVLTIISLIILI"
     CDS             5594..5617
                     /codon_start=1
                     /label=gBlock replacing part
                     /translation="LWQKKPR"
     CDS             5618..5817
                     /codon_start=1
                     /label=Stargazin 203-319 targeting S->D
                     /translation="RHKQLRATARATDYLQADAITRIPDYRYRYQRRDRDDDRDTEPDH
                     DRDADPVGVKGFNTLPDTEID"
     primer_bind     complement(5806..5865)
                     /label=Fragment.REV
     CDS             5818..5819
                     /codon_start=1
                     /label=PDZ domain
                     /translation=""
     protein_bind    complement(5949..5996)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     intron          6018..6150
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(6172..6619)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   6663..6827
                     /label=ITR
     primer_bind     complement(6868..6884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6892..6908
                     /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(6916..6946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6961..6982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgaatt aggttaatta aggctgcgcg ctcgctcgct
     2581 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     2641 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
     2701 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tccttatcga
     2761 ttttaccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacatc tccccctgaa
     2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2941 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgctcga agcggccgcc
     3001 cgggtcgagg tcgacggtat cgatgcgggg aggcggccca aagggagatc cgactcgtct
     3061 gagggcgaag gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa
     3121 ggtccgctgg attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg
     3181 tggcaacaca ggcgagcagc caaggaaagg acgatgattt ccccgacaac accacggaat
     3241 tgtcagtgcc caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt
     3301 ccgccgtggc aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg
     3361 caatgcccca accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc
     3421 tgacaacggg ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga
     3481 aaatgaaagc catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac
     3541 atagcgtaaa aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat
     3601 ccagaggttg attatcgata agcttcctgc agggaagttc ctattccgaa gttcctattc
     3661 tgaactaagt ataggaactt cttggatcaa aataggagcg atcgcaatgg aagttcctat
     3721 tccgaagttc ctattctcta gaaagtatag gaacttcgtt taaactggcc ggccactagt
     3781 agatctgaat tcgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
     3841 gttccaggtt ccactggtga cggatccgcc gccggaagta ctcttgataa gatcgctaaa
     3901 aatggtgtaa ttgttgtcgg ccaccgagac tcttctgtgc cattcagcta ttatgattct
     3961 caacacaaag tggtgggcta ttcacaagag tattctaacg ccattgttga agcagttaag
     4021 aagaagttga ataaaccaga ccttcaagta aaactcattc ctataacatc ccaaaaccgc
     4081 atacctctct tgcaaaacgg cacttacgat ttcgagtgtg ggagcactac taataatgtc
     4141 gaacgccaga aacaagccgc attctccgac accatcttcg tggtgggcac tcgactgttg
     4201 acaaagaaag gcggtgatat taaggatttc gcaaatctga aggataaggc agtcgtggtc
     4261 accagcggga ccacctcaga agtactcctt aataaactca atgaagaaca aaagatgaat
     4321 atgcgaatca tctctgctaa ggaccacggt gactcttttc gcacattgga aagtgggcgc
     4381 gccgtagcat ttatgatgga cgactctctg ctcgctggcg agcgcgccaa agccaaaaaa
     4441 ccagacgact gggaaattgt aggtaaacct caaagccaag aggcatatgg ctgcatgttg
     4501 agaaaagatg acccccaatt caagaaactg atggacgaca ctatagctca ggtccagact
     4561 tcaggggaag ctgagaagtg gtttgataaa tggttcaaga acccaatctt ggtaaatcat
     4621 accgtttata ttaccgctga caaacaaaag aatgggataa aggcaaattt caagattcac
     4681 cataacgtag aggatggaag cgtgcagttg gcagaccatt atcaacagaa tacccctatt
     4741 ggcgacggac ccgtgctcct ccctgacaac cactacttgt cctatcaaag tgttctctca
     4801 aaggatccca atgagaagcg agaccacatg gtacttctcg aattcgttac cgccaccggg
     4861 aattcacttg gcatggatga attgtataag gggggtactg gcggcagcat gtccaagggt
     4921 gaggagcttt ttaccggagt agtaccaatt ctcgttgaac ttgacggaga cgtcaacggg
     4981 cacaagttta gcgtacgagg tgagggggag ggtgatgcaa ccaacggtaa gcttactctc
     5041 aaattcatat gcactaccgg aaagctgcca gtaccttggc caacactggt cactaccctc
     5101 tcctatggcg ttcaatgttt tgctcgatac cctgaccaca tgaaacagca tgacttcttt
     5161 aaatcagcca tgcctgaagg gtatgtccag gagcggacta tatcatttaa agacgatgga
     5221 acctataaga ctcgggctga agtgaagctc gagggtgaca cattggttaa tcggatcgag
     5281 cttaagggca tagattttaa agaagatggt aatattcttg gccacaaatt ggagtacaat
     5341 tttaataatc ctttgaacat gaactttgag ctctccgatg aaatgaaggc actctttaaa
     5401 gagccaaacg acaaagctct gaaactgcag gtcgacgaac aaaaactcat ctcagaagag
     5461 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     5521 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     5581 atcatcctca tcatgctttg gcagaagaag ccacgtgata gacacaagca gttgcgagct
     5641 acagcacgcg ccaccgatta tctgcaagct gatgcaataa ctagaatccc tgattataga
     5701 tacagatatc aacgaaggga tcgcgatgat gatcgagata cagagccaga tcatgatcgg
     5761 gatgctgatc ccgtcggggt gaaaggtttc aacactcttc cagatacaga gatagataat
     5821 ccccagacga cttcagtgta gtaaaccggt acgcgtttaa ttaagtttaa acgaagttcc
     5881 tatacttagt tcagaatagg aacttcggaa taggaacttc agagcgcttt ggccgctccg
     5941 gccaaaggga agttcctata ctttctagag aataggaact tcggaatagg aacttcggcg
     6001 cgccggatcc ttatcacctg tggagagaaa ggcaaagtgg atgtcagtaa gaccaatagg
     6061 tgcctatcag aaacgcaaga gtcttctctg tctcgacaag cccagtttct attggtctcc
     6121 ttaaacctgt cttgtaacct tgatacttac ctgctagcag cttgaattcg actgcgctct
     6181 caggcacgac acgactcctc cgctgcccac cgcagactga ggcagcgctg agtcgccggc
     6241 gccgcagcgc agatggtcgc gcccgtgccc ccctatctcg cgcctcgcgt ggtgcggtcc
     6301 ggctgggccg gcggcggcgc ggacgcgacc aaggtggccg ggaaggggag tttgcggggg
     6361 accggcgagt gacgtcagcg cgccttcagt gctgaggcgg cggtggcgcg cgccgccagg
     6421 cgggggcgaa ggcactgtcc gcggtgctga agctggcagt gcgcacgcgc ctcgccgcat
     6481 cctgtttccc ctccccctct ctgatagggg atgcgcaatt tggggaatgg gggttgggtg
     6541 cttgtccagt gggtcggggt cggtcgtcag gtaggcaccc ccaccccgcc tcatcctggt
     6601 cctaaaaccc acttgcactc atacgcaggg ccctctgcag agatcttcct agagcatggc
     6661 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     6721 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     6781 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag cctaattaag
     6841 gccttaatta aatctggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6901 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6961 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7021 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7081 tgggcgct
//
