LOCUS       sequence-326672-        6701 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV promoter driving GFP labeled version of HyPep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6701)
  AUTHORS   Hawk JD, Wisdom EM, Sengupta T, Kashlan ZD, Colon-Ramos DA
  TITLE     A genetically encoded tool for reconstituting synthetic modulatory 
            neurotransmission and reconnect neural circuits in vivo.
  JOURNAL   Nat Commun. 2021 Aug 9;12(1):4795. doi: 10.1038/s41467-021-24690-9.
  PUBMED    34373460
REFERENCE   2  (bases 1 to 6701)
  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: "Nat 
            Commun."; date: "2021-08-9"; volume: "12(1):4795. doi"; pages: " 
            10.1038/s41467-021-24690-9"
COMMENT     Sequence Label: pJH119_pCMV_HyPep-emGFP
FEATURES             Location/Qualifiers
     source          1..6701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     874..893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        874..892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    961..985
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1465..1467,1468..2181)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1507..1528)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1768..1787)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2115..2136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2212..2236)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2265..2283)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2266..2283)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2303..2320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2309..2533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2579..3007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2666..2685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2876..2897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3016..3036)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3021..3350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3201..3336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3263..3282
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3417..4211
                     /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(3471..3490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4081..4100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4385..4506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4422..4441)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4476..4495
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4555..4571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4555..4571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4568..4590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4579..4595
                     /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(4603..4609,4610..4627,4628..4633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4648..4669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4786..4803)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4957..5545)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5037..5056)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5716..6507,6508..6576))
                     /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     6339..6358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6577..6681)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      901 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attgattaat
      961 acaagtttgt acaaaaaagc aggcttagcc accatgctgg gaaacaaaag aatgggactg
     1021 tgtggactga ccctggccct gtctctgctg gtgtgtctgg gaattttggc tgagggatat
     1081 ccatctccag ataatccagg agaggacgcc ccagctgaag atgagacagg agctgaagat
     1141 ctgggagagg agcacgcttg gaaggctagc catagacaga aaagacagtg gctgggagga
     1201 agattcggaa aaagatctgg agaagaggag gatcatgata tgatgaagag acagtggctc
     1261 ggaggaagat ttggaaagag aagcggagat gaggaggccg agactagtca cagacagaag
     1321 agacagtggt ttaacggaag attcggcaaa agatctggcg aggaggaaga tcacgatatg
     1381 atgaaaagac agtggttcaa cggaagattt ggcaaaagat ccggagatga ggaagcctct
     1441 agaggatccc acagacaaaa aagagtgagc aagggcgagg agctgttcac cggggtggtg
     1501 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1561 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1621 ctgcccgtgc cctggcccac cctcgtgacc accttgacct acggcgtgca gtgcttcgcc
     1681 cgctaccccg accacatgaa acagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1741 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1801 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1861 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa ggtctatatc
     1921 accgccgaca agcagaagaa cggcatcaag gtgaacttca agacccgcca caacatcgag
     1981 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2041 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2101 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2161 atggacgagc tgtacaagta aaagagatct ggagatgaag aatgatcatg aacccagctt
     2221 tcttgtacaa agtggtcctc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     2281 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2341 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2401 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2461 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2521 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2581 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2641 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2701 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2761 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2821 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2881 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2941 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3001 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3181 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3241 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3301 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3361 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3421 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3481 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3541 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3601 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3661 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3721 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3781 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3841 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3901 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3961 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4021 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4081 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4141 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4201 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4261 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4321 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4381 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4441 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4501 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4561 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4621 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4741 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4801 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4861 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5461 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5521 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5581 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5641 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5701 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5761 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5821 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5881 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5941 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6001 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6061 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6121 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6181 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6241 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6301 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6361 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6421 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6481 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6541 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6601 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6661 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg c
//
