LOCUS       sequence-316174-        6857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV promoter driving HyCal component (HyNaC9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6857)
  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 6857)
  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: pJH75_pCMV_hyCal9
FEATURES             Location/Qualifiers
     source          1..6857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    716..740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2123..2147)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2176..2194)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2177..2194)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2214..2231)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2220..2444
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2490..2918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2577..2596)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2787..2808
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2927..2947)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2932..3261
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3112..3247
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3174..3193
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3328..4122
                     /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(3382..3401)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3992..4011
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4296..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4333..4352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4387..4406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4466..4482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4466..4482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4479..4501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4490..4506
                     /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(4514..4520,4521..4538,4539..4544))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4559..4580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4697..4714)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4868..5456)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4948..4967)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5627..6418,6419..6487))
                     /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     6250..6269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6488..6592)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6667..6686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattga ttaatacaag
      721 tttgtacaaa aaagcaggct tagctagcgc caccatgagc gaggaagatg agaagaaaat
      781 taagagccca catgaacaga gaaatgatag agtgaaagaa cacgtggctc atctgattaa
      841 gaatgtgagc ttccatggac tgagctatgt ggctgataaa agaaacaact acttcagaag
      901 agctatttgg ttcctgatca ccgtgggagc cttcatctat gctgtggaaa aggtgtatga
      961 gtctaccgtg aattacttta gctatccatt caagacagct agaatgaaaa tctatgtgaa
     1021 tgaactgaac ttcccagctg tgagcttctg caatctgaac gattttctgt tttctaagct
     1081 gaatggaacc aaactggatg aaagcatcct gtacccagat gatccagaga aaaacaatgt
     1141 gtctgaaatt gagatttcca atatcacatc tgacgctaca atcagactgg atcagatgct
     1201 ggtggattgc gaattcgagg gaaaaaagtg tacacacgag aatttcaccg atttctggac
     1261 catgcaggga gagctgtgct tcacattcaa tagcggaaaa aattctcact ctctgctgaa
     1321 ggtgtctgga gtgggagtgc tgagatctct gaagctgacc attaacgtgc agcattatga
     1381 gtattacaga gatgagatgg ccggaggaat tcacctgatg attcatggac aggatgaaga
     1441 gccagtgaaa atgcagggac agatcgtgtc tccaggatat tctttctatg tgaaggtgga
     1501 aaagaagacc atcatgaacc tggagaaacc atataagaca gagtgcggaa ccgtgaaact
     1561 gaaatatttt gatagatact ccatgcacac atgctggctg gagcagctga ccgattatgt
     1621 gaataagatg tgccactgca aggacttctt catgccagga aatattccat attgttctct
     1681 gccagaactg cagaactgta cctggattga gtgggctaaa ttcaataagg ataaaatgta
     1741 caagtgccca ctgccatgta agattgacct gtacggagtg tctctgtcta gagccctgtt
     1801 cccaaccaca cagtatagct ctatcctggc cgagcagttc agaaaacagc cacatgtgct
     1861 gtctatcgtg cacaacatta cagatgagct gctgttcatg agagataacc tgctgagatt
     1921 cattatctat tacgatgatc tgagctatga ggtgctggaa cagaaaccat cttatgagac
     1981 actggtgtgg ctgggagaca ttggaggaca gatcggactg tttatcggag ccggagtgat
     2041 gagctacttc gagttcctgg attgcctggc catcgtgatc tacaccagat ttttccagaa
     2101 gttcacaagc agctgaggat ccacccagct ttcttgtaca aagtggtcct cgagcatgca
     2161 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2221 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2281 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2341 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2401 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2461 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2521 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2581 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2641 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2701 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2761 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2821 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2881 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2941 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3001 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3061 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3181 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3241 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3301 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3361 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3421 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3481 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3541 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3601 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3661 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3721 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3781 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3841 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3901 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3961 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4021 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4081 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4141 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4201 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4261 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4441 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4621 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4681 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4741 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4801 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4861 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4921 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4981 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5041 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5101 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5161 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5221 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5281 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5341 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5401 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5461 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5521 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6601 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6661 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6721 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6781 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6841 ggccagatat acgcgtt
//
