LOCUS       Exported                5727 bp DNA     circular SYN 25-APR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5727)
  AUTHORS   Yan Zhang
  TITLE     Direct Submission
  JOURNAL   Exported May 4, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5727
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     primer_bind     complement(270..281)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     331..342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(794..805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1482..2728
                     /codon_start=1
                     /label=jGCaMP8s (var 688)
                     /translation="RRKKTFKEVATAVKIIAMLMGLKINVYIKADKQKNGIKANFHIRH
                     NIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLSVESKLSKDPNEKRDHMVLLEFVTAAG
                     ITLGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEYKEA
                     FSLFDKDGDGTITTKELGTVMRSLGHNPTEAELQDMINEVDADGDGTIDFPEFLTMMAR
                     KMKYRDTEEEIREAFGVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGD
                     GQVNYEEFVQMMTAK"
     polyA_signal    2752..2873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2763..2782)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2817..2836
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3035..3164
                     /label=AAV2 ITR
     primer_bind     complement(3081..3092)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3142..3153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3183..3200)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3183..3199)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3192..3214)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3341..3796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3428..3447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3442..3457)
                     /label=CP864
     primer_bind     3638..3659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3927..4787
                     /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"
     primer_bind     complement(4145..4164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4958..5546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5447..5466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5491..5506
                     /label=CP866
     primer_bind     5700..5717
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagaattc gccaccatgg gccatcatca ccatcatcac acgcgtcgca agaagacctt
     1501 caaggaggtg gccaccgccg tgaagatcat cgccatgctg atgggcctga agatcaacgt
     1561 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa
     1621 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1681 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg gagtccaaac tttcgaaaga
     1741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1801 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     1861 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1921 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     1981 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2041 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2101 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     2161 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2221 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     2281 gccggaccaa ctgactgaag agcagatcgc agaatacaaa gaggctttct ccctatttga
     2341 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctcgg
     2401 acacaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga
     2461 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga
     2521 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat
     2581 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga
     2641 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga
     2701 gtttgtacaa atgatgacag cgaagtaagc ggccgcttcg agcagacatg ataagataca
     2761 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2821 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2881 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     2941 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta
     3001 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc
     3061 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3121 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa ttaacctaat
     3181 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3241 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3301 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3361 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3421 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3481 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3541 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3601 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3661 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3721 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3781 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4021 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4081 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4141 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4441 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4621 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4801 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5101 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5641 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5701 gcgagtcagt gagcgaggaa gcggaag
//
