LOCUS       Exported                6056 bp ds-DNA     circular SYN 16-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Chapman lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 17, 2022 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6056
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           1076..1118
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     misc_feature    1127..1216
                     /label=Bovine pre-prolactin
     CDS             1235..1945
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    1946..1987
                     /label=Linker
     CDS             1988..3145
                     /codon_start=1
                     /label=Syt9_M
                     /translation="FPEPPTLGSPAPKTPPDSSRIRQGAVPAWVLATIVLGSGLLVFSS
                     CFCLYRKRCRRRMGKKSQAQAQVHLQEVKELGRSYIDKVQPEIEELDRSPSMPGQQVSD
                     KHQLGRLQYSLDYDFQTGQLLVGILQAQGLAALDLGGSSDPYVSVYLLPDKRRRHETKV
                     HRQTLNPHFGETFAFKVPYVELGGRVLVMAVYDFDRFSRNDAIGEVRVPMSSVNLGRPV
                     QAWRELQVAPKEEQEKLGDICFSLRYVPTAGKLTVIVLEAKNLKKMDVGGLSDPYVKVH
                     LLQGGKKVRKKKTTIKKNTLNPYYNEAFSFEVPCDQVQKVQVELTVLDYDKLGKNEAIG
                     RVAVGAAVGGAGLRHWADMLANPRRPIAQWHSLRPPDRARPIPAP"
     polyA_signal    3170..3291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3472..3927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4259..4363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4364..5224
                     /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"
     rep_origin      5395..5983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt acttaatacg actcactata ggctagcctc gagaattcga tatcgcttgt
     1081 tctttttgca gaagctcaga ataaacgctc aactttggca gataccatgg acagcaaagg
     1141 ttcgtcgcag aaagggtccc gcctgctcct gctgctggtg gtgtcaaatc tactcttgtg
     1201 ccagggtgtg gtctccgata tcggtaccac gcgtagtaaa ggagaagaac ttttcactgg
     1261 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag
     1321 tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac
     1381 tggaaaacta cctgttccat ggccaacact tgtcactact ttaacttatg gtgttcaatg
     1441 cttttcaaga tacccagatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     1501 aggttatgta caggaaagaa ctatattttt caaagatgac gggaactaca agacacgtgc
     1561 tgaagtcaag tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt
     1621 taaagaagat ggaaacattc ttggacacaa attggaatac aactataacg atcaccaggt
     1681 gtacatcatg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttagacacaa
     1741 cattgaagat ggaggcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     1801 tgggcccgtc cttttaccag acaaccatta cctgtttaca acttctactc tttcgaaaga
     1861 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     1921 acatggcatg gatgaactat acaaagaatt ctcaggcgga agcggaggct ccggcgggag
     1981 cggcggattc ccggaacccc cgaccctggg gtctccagcg cctaagacac ctccagactc
     2041 cagccgcatc aggcagggcg cagtgcctgc ctgggtcctg gctaccatcg tgctgggctc
     2101 gggcctcctg gtcttcagca gttgtttctg tctctaccgg aagcgctgtc ggagacggat
     2161 gggcaaaaag agtcaggccc aagcccaagt ccatcttcag gaagtgaagg agctgggccg
     2221 gagttacata gataaggttc agcctgaaat agaggagctg gaccggtcac catccatgcc
     2281 tggacagcag gtatcagaca aacaccagct aggccgactg cagtattcac tggactatga
     2341 cttccagact ggccagctcc tggtgggcat cttgcaagct cagggactgg ctgcgttgga
     2401 cctaggaggt tcctcagatc cctatgttag tgtctatctg ctgccagaca agcggaggcg
     2461 acatgagacc aaggtgcatc ggcagaccct gaatccacac tttggagaga catttgcctt
     2521 caaggtccca tacgtggaac tagggggcag agtgctggtc atggcggtgt atgactttga
     2581 tcggttctcc cgcaacgatg ccatcgggga ggtgcgggtg cctatgagtt cagtgaacct
     2641 ggggcggccg gtgcaggcct ggagagagct gcaggtggct cccaaagagg agcaagagaa
     2701 actgggggac atctgcttct ctctccgtta tgtccccacg gccgggaagc tcaccgtcat
     2761 tgtcttggaa gctaaaaacc tgaagaaaat ggatgtagga ggactctcag atccctatgt
     2821 gaaggtgcac ctgctccagg gaggtaaaaa ggttcgcaag aagaaaacca ccattaagaa
     2881 gaacaccctg aacccctatt acaacgaggc cttcagcttt gaggtgccct gtgaccaggt
     2941 gcagaaagtc caggtggagc tgactgttct ggattatgac aaactgggga agaatgaggc
     3001 catcgggaga gtggcggtgg gcgcagcagt cggtggggct ggcctgcggc actgggctga
     3061 catgttggcc aaccccaggc gccccattgc ccagtggcac tctctgcggc cccctgacag
     3121 agccaggcca ataccagcac cctgagcggc cgcttcgagc agacatgatt aagatacatt
     3181 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3241 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3301 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
     3361 taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa tagcgaagag
     3421 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     3481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3661 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3781 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     3841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3901 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3961 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4021 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4081 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4141 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4201 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac agatct
//
