LOCUS       sequence-456530-        6378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6378)
  AUTHORS   Bailes HJ, Zhuang LY, Lucas RJ
  TITLE     Reproducible and sustained regulation of Galphas signalling using a 
            metazoan opsin as an optogenetic tool.
  JOURNAL   PLoS One. 2012;7(1):e30774. Epub 2012 Jan 24.
  PUBMED    22292038
REFERENCE   2  (bases 1 to 6378)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2012"; volume: "7(1)"; pages: "e30774. Epub 2012 Jan 24"
COMMENT     Sequence Label: pcDNA3.1 JellyOp
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(1969..1986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1975..2199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2245..2673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2332..2351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2542..2563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2682..2702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2687..3016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2867..3002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2929..2948
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3083..3877
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3137..3156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3747..3766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4051..4172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4088..4107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4142..4161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4221..4237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4221..4237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4234..4256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4245..4261
                     /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(join(4269..4275,4276..4293,4294..4299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4314..4335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4452..4469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4623..5211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4703..4722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5382..6173,6174..6242))
                     /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     6005..6024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6243..6347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa gttcgaatgc atctagatcc aataacttaa gcttcaccat
      901 gggagcgaat ataacagaaa tattgtcagg ctttttagct tgtgtagtat ttctctctat
      961 aagcttgaat atgatagttc taattacatt ttaccgctta cgacataaat tggcttttaa
     1021 agatgctctc atggctagta tggcgttcag tgatgttgta caagctatcg taggatatcc
     1081 tttagaggta ttcactgtag tagacggaaa atggacattc ggaatggaat tatgtcaagt
     1141 tgcaggattt ttcattactg cccttggtca agtttccatc gctcatctca ctgctctcgc
     1201 cttagatcga tacttcacag tgtgcagacc tttcgtggca actgcgattc acggatcaat
     1261 gagaaatgca ggtatggtca tatttgtttg ctggttctac gcgtccttct gggcagtcct
     1321 acccttagtt ggatggtcaa attatgacgt ggagggtgac ggtatgcgat gctccatcaa
     1381 ctgggcagac gacagtccta aaagctactc atacagggtt tgtttattcg tattcatcta
     1441 cctgattcct gttcttttga tggtggctac ttacgtgttg gttcaaggag agatgaagaa
     1501 tatgcgaggt cgtgcagcac agttgtttgg ttcagaatcc gaggctgcac taaagaatat
     1561 caaggccgaa aagcgacaca caaggctggt tttcgtcatg atcctttctt tcatcgtagc
     1621 ttggacccca tataccttcg tcgccatgtg ggtttccttt ttcacgaaac aacttgggcc
     1681 aataccttta tacgttgata ctttggcagc tatgcttgca aagtcgtcgg ctatgttcaa
     1741 ccccatcatt tactgcttcc tccacaagca attcagaaga gctgtattac gtggtgtttg
     1801 tggaagaatt gtcggtggta atgccattgc tccttccagt acggctgttg aacctggaca
     1861 aactttggct tcaggaacag ccgaatctgc tagcacggag acgagccagg tggccccggc
     1921 ctaagcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     1981 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2041 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2101 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2161 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2221 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2281 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2341 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2401 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2461 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2521 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2581 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2641 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2701 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2761 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2821 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2881 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2941 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3001 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3061 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3121 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3181 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3241 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3301 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3361 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3421 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3481 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3541 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3601 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3661 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3721 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3781 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3841 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3901 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3961 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4021 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4081 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4141 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4201 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6361 aaagtgccac ctgacgtc
//
