LOCUS       Exported                6963 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    ElectricPk
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  AUTHORS   Barnett L, Platisa J, Popovic M, Pieribone VA, Hughes T
  TITLE     A fluorescent, genetically-encoded voltage probe capable of 
            resolving action potentials.
  JOURNAL   PLoS One. 2012;7(9):e43454. Epub 2012 Sep 6.
  PUBMED    22970127
REFERENCE   2  (bases 1 to 6963)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6963
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     11..30
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11..29
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1011..1032
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1138..1159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1399..1418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1536..1553)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1542..1766
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1812..2240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1899..1918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2109..2130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2249..2269)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2254..2583
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2434..2569
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2496..2515
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2632..3657
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    3787..3908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3824..3843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3878..3897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3957..3973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3957..3973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3970..3992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3981..3997
                     /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(4005..4035)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4050..4071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4188..4205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4359..4944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4439..4458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5115..5975)
                     /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     5738..5757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5976..6080)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6155..6174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6346..6725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6726..6929
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6880..6900
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttccatggag
       61 ggattcgacg gttcagattt tagtcctcca gctgatttag ttggcgttga cggtgcagtc
      121 atgcggaacg tcgttgacgt cacgataaat ggtgacgtca ctgctccgcc gaaagctgcg
      181 ccaagaaaaa gtgaatcggt aaagaaagtt cattggaatg atgtagacca aggaccgagt
      241 gaaaaaccag agacaagaca ggaggaacga atagatatac ccgagatttc aggtctatgg
      301 tggggcgaga atgaacatgg agtggacgat gggagaatgg agatacctac tactggtgta
      361 ggtcgcgtcc agtttcgtgt ccgagcagtg attgatcatc tagggatgcg agtctttgga
      421 gtcttcctaa ttttcttgga catcatcctc atgatcattg atctcagtct tccaggaaaa
      481 agtgaatctt cacaatcctt ttatgacggg atggctttgg ctctttcttg ttatttcatg
      541 ctggatttag gattaaggat atttgcctac gggcccaaga atttcttcac caacccctgg
      601 gaggttgctg atggtttgat tatcgttgtc acattcgtcg tcacgatatt ttacactgtg
      661 ttagatgaat acgtgcaaga aacaggagcc gattgtttgg ggcagttggt tgtgttggcc
      721 cgtttgctgc gtgtggttcg attagcaaga atattttatt cccaccaaca aatgaaggct
      781 tcaatgagtg tgattaaaag tcataacgtc tatatcaagg ccgacaagca gaagaacggc
      841 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac
      901 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      961 ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg
     1021 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
     1081 accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1141 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc
     1201 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1261 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1321 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag
     1381 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1441 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1501 atcctggggc acaagctgga gtacaactag cgcgccctcg actgtgcctt ctagttgcca
     1561 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1621 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1681 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1741 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1801 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1861 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1921 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1981 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2041 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2101 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2161 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2221 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2461 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2521 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2581 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gatgaaaaag
     2641 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     2701 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
     2761 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
     2821 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
     2881 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
     2941 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
     3001 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
     3061 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
     3121 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
     3181 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
     3241 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
     3301 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
     3361 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
     3421 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
     3481 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
     3541 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
     3601 gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
     3661 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3721 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3781 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3841 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3901 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3961 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4021 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4081 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4141 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4201 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4261 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4321 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4381 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4441 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4501 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4561 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4621 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4681 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4741 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4801 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4861 tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt
     4921 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4981 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5041 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5101 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5161 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5221 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5281 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5341 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5401 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5461 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5521 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5581 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5641 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5701 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5761 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5821 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5881 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5941 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6001 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6061 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6121 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6181 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6241 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     6301 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     6361 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6421 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6481 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6721 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6781 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6841 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6901 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     6961 ggc
//
