LOCUS       sequence-316173-        7010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV promoter driving HyCal component (HyNaC7).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7010)
  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 7010)
  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: pJH74_pCMV_hyCal7
FEATURES             Location/Qualifiers
     source          1..7010
                     /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(2276..2300)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2329..2347)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2330..2347)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2367..2384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2373..2597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2643..3071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2730..2749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2940..2961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3080..3100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3085..3414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3265..3400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3327..3346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3481..4275
                     /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(3535..3554)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4145..4164
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4449..4570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4486..4505)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4540..4559
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4619..4635)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4619..4635)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4632..4654)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4643..4659
                     /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(4667..4673,4674..4691,4692..4697))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4712..4733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4850..4867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5021..5609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5101..5120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5780..6571,6572..6640))
                     /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     6403..6422
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6641..6745)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6820..6839)
                     /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 caccatgaga atcagactga aggagagaat
      781 gagaaaaatt gtgcaggaaa gactgaccgt gaaacagatt tttaaaagat atatcgagag
      841 ctctaccctg cacggatttt gttatgtgtg tatggacaca ttcctgggaa gaagactgat
      901 ttgggccgtg ctgatgatcc tgggagccat ctatttcatt tttaaactga gatacggcat
      961 caaggaatac tttgactatc cattcagcac cctgtctaca gtggaatatg tggacgatct
     1021 gctgtttcca gccgtgtctg tgtgcgctac caatagctac attgcctctc aggtgtatac
     1081 caatcagctg aatacaatgt ataaggaggg aagactgcca ctggataaca accagtctat
     1141 cccagaatat aacattccag gagacgagct ggtgaagacc ctgaagaaca gcagcctgac
     1201 aattgagtct ctgctgaagt attgtgactg gatcatgcag gacacatctc acccactggt
     1261 gaccccaaat aattgtggag ctctgaattt tacaagctat tttaattaca agggagagca
     1321 gtgccatacc ctgaacagcg gagctaaagg acatgaactg ctgaaggtga gcgatgtggg
     1381 aatctctcat ggatacgaac tggtgtttga cctgcagaca aacgaggtta ttaagaacta
     1441 tcagctgagc ggaatgagaa tcgtgatcca tgatcaggtg ttcccaccac agctggtgga
     1501 tggattcttt atcagcccag gattcaagac ctacatcaag ctgggaatta cccagtctca
     1561 gtctctgcca ccaccatatt ctacagaatg cggacagaaa aaactgaagt attatgctat
     1621 ctatagccag agactgtgcc tgctggagac cctgacagac ttcacaggag acctgtgcgg
     1681 atgtagagat gtgtttatgc cagagaacgg actgccattc tgtagcctgc aggaactgta
     1741 tagctgcatg tacccagcta aagagtcttt tagcgaattc accatgagaa aagagtgtcc
     1801 aagcgattgc gaagaaagaa cctttagcta cgagctgtct gaggctagat atctgcacaa
     1861 tccaccaatt ggactgtctc tgagcagact ggataaactg cagctggaca agtctctgcc
     1921 aagcgaggct cacctgtcta aactggctaa aagcctgaca ccaaaagaac tggatgccta
     1981 catcgaaagc aacatcatca gcgtgattct gtttttcgga gataccagaa tcgattacca
     2041 cgaacaggaa gccacaaacg actttttcca gttcctggga aatatgggag gagaatttgg
     2101 actgatgctg ggagcttccc tgctgacctt cgtggaattc attgacctgt ttattgtgct
     2161 gctgtaccac cagatgctga gactgtataa cctgagaaag attccagaca tttttgccag
     2221 caaaagaaac agaaataagg aaaaaaccaa gaagtgtgag aacgtgtgag gatccaccca
     2281 gctttcttgt acaaagtggt cctcgagcat gcatctagag ggccctattc tatagtgtca
     2341 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2401 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2461 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2521 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2581 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2641 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2701 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2761 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2821 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2881 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2941 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3001 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3061 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3181 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3241 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3301 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3361 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3421 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3481 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3541 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3601 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3661 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3721 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3781 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3841 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3901 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3961 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4021 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4081 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4141 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4201 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4261 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4321 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4381 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4441 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4501 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4561 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4621 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4681 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4861 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4921 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4981 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5041 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5101 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5161 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5221 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5281 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5341 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5401 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5461 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5521 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5581 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5641 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5701 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5761 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5821 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5881 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5941 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6001 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6061 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6121 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6181 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6241 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6301 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6361 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6421 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6481 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6541 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6601 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6661 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6721 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     6781 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     6841 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     6901 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     6961 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
