LOCUS       sequence-468387-        7400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes oxytocin/vasopression like receptor 2 in H.exemplaris, 
            untagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7400)
  TITLE     Gruber lab plasmids 2025
REFERENCE   2  (bases 1 to 7400)
  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"
COMMENT     Sequence Label: Tardigrada_OTR-like2_untagged
FEATURES             Location/Qualifiers
     source          1..7400
                     /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     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      933..942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2243..2245,2246..2959)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2243..2956
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2285..2306)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2546..2565)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2893..2914
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2991..3008)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2997..3221
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3267..3695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3354..3373)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3564..3585
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3704..3724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3709..4038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3889..4024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3951..3970
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4105..4899
                     /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(4159..4178)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4769..4788
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5073..5194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5110..5129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5164..5183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5243..5259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5243..5259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5256..5278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5267..5283
                     /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(5291..5297,5298..5315,5316..5321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5336..5357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5474..5491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5645..6233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5725..5744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6404..7195,7196..7264))
                     /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     7027..7046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7265..7369)
                     /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 attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatggcg gcagcgttta acgtgtcatc
      961 atcaacttta tcgccttcgt ccgggcagca gccgttcctg aactacacgc cgtttgagga
     1021 caaacgcgac gaggacctcg ccaaagtgga gattggaata ctggggacgg tgttcgcgct
     1081 cgctctgctg ggcaatgtcg cggtgctgtc ggtcctgctg tggcggagaa agaaggtcac
     1141 acggatgtat tttttcatct tccacctctc cctctctgat ctcgtcgtgg cattcttcaa
     1201 catatttccg caaatggtgt gggacatcac ttggcggttt caggggaatg atcttggctg
     1261 caaaatcgtc aaattcctgc aggtttttgc tctttacctg tcaacatggg tcctcgttat
     1321 gatggccatt gaccggtacc aggcgatctg tcaacccctc aatgcgtaca actggcgcgc
     1381 taaacgctgc accatgatga tcgcatatgc ctggattctg tcgggacttt gtagtctgcc
     1441 gcaaattttc atattctccc agatggaaat catggaaggc agcggggtct acgactgctg
     1501 ggcgaccttt atcgaacctt ggggctcaaa agcttatgtg atttggtttg ccggatctat
     1561 ttatataata ccgctggtga tactatgcgt gacctacggc tacgtgtcct ggacaatctg
     1621 gcagaccatg cggcaaaaac aaagcggtag tctgagagtg aaaaatcgac ctctcggatc
     1681 gaccacagga agtggccata acctgctacc ggattctcgg acacattcga cctacaaacg
     1741 cggcctgagt ttcacgcgag ccaagttacg gacagtgaaa ttgacgcttg tagtgattat
     1801 tgcctatctg atttgttgga cacctttcgt tgtggctctg ctctggacta cctttttccc
     1861 ggagaccgct tatctcagcg agccatccct tgtcatcctt ttattgttgg cgaatctcaa
     1921 cagctgttcg aatccatgga tttacttgtt cttctccctc tctcgctact ttcgatgcgt
     1981 ggagtggtgc ttggaccggc tggaaagcga agagcaaaag cgactgaggg ctgtgaccca
     2041 gaccgtctgt ctgccgcagt tcggggccag cagctccgcc agctcctggt acatccacaa
     2101 caagggaccc aacaacaccg ccggcggagg tggcgcgagt ggtttgccct tgaccgcctc
     2161 ggtccaggct aacggccacg gcggctatgt ctgcaaactt ttaccgcgct caggcaccac
     2221 tcacgatcta tgaggcggcc gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2281 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2341 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2401 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2461 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2521 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2581 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2641 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2701 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2761 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2821 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2881 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctcacggcat
     2941 ggacgagctg tacaagtaat ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3001 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3061 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3121 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3181 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3241 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3301 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3361 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3421 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3481 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3541 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3601 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3661 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3721 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3781 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3901 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3961 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4021 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4081 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4141 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4201 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4261 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4321 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4381 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4441 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4501 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4561 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4621 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4681 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4741 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4801 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4861 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4921 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5041 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5221 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5821 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6061 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7381 gaaaagtgcc acctgacgtc
//
