LOCUS       sequence-413050-        5856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes the inotocin receptor (oxytocin/vasopressin-like) from the 
            ant Lasius niger, tagged with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5856)
  AUTHORS   Di Giglio MG, Muttenthaler M, Harpsoe K, Liutkeviciute Z, Keov P, 
            Eder T, Rattei T, Arrowsmith S, Wray S, Marek A, Elbert T, Alewood 
            PF, Gloriam DE, Gruber CW
  TITLE     Development of a human vasopressin V(1a)-receptor antagonist from an
            evolutionary-related insect neuropeptide.
  JOURNAL   Sci Rep. 2017 Feb 1;7:41002. doi: 10.1038/srep41002.
  PUBMED    28145450
REFERENCE   2  (bases 1 to 5856)
  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: "Sci Rep."; 
            date: "2017-02-1"; volume: "7:41002. doi"; pages: " 
            10.1038/srep41002"
COMMENT     Sequence Label: Lnig-INTR-GFP
FEATURES             Location/Qualifiers
     source          1..5856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1796..1805
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1802..1804,1805..1807,1808..2521)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1802..1804,1805..1807,1808..2518)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1847..1868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2108..2127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2455..2476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2644..2765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2681..2700)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2735..2754
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2772..3227)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2909..2930)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3121..3140
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3254..3358
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3360..3717
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3360..3551
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3383..3403)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3568..3703
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3752..4546
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3752..4546
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3806..3825)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4416..4435
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4734..4753)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4778..4825
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5154..5742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5643..5662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcatgtcg tatgactcga atacgtcgtt
      661 atcttcgcca tcgtcgttgt cgtcttcgat ggagacgccg gacgacacga gggacgagta
      721 tctcgccaga tgggaaatcg cggttttaac tatcatattt ctcgtcacgc tcatcggcaa
      781 caccctgatc ctgtttgccc tttacgccag gcgacgttat cagcgtcgaa agttcaccag
      841 gatgtatttc ttcattctac atttgagcat cgccgacttg ttgacgggtt tgctcgatgt
      901 tttaccgcaa cttgcttggg atataacatt cagatttcag ggcggagcgg tgctctgcaa
      961 actggtcaaa ttcggacagc cgttcggtgt gtatctcagt tcctatgtcc tgacagttac
     1021 cgcgatggac aggtattacg ccatttgcca tcccttcctt tactgcagca tcacgtctag
     1081 aagatcgaaa atgatggtgt atggagcatg gacgctcgcc gctatacttt gtgtaccgca
     1141 ggtttttata ttctcatacc aggaaatatc gccaggcgtt tgggaatgtt gggccacctt
     1201 ttacctgaaa tatggcgaga gagcttatgt cacttggtac agtatcatgc agttcctatt
     1261 accctttata gtacttgtat acacgtacac aaagatatgt atagcaatat ggacgagcag
     1321 taagatgtca ggagtcgttg atctcaaaaa gaacaacaag gctaatttct cgcagcgaaa
     1381 tcgggagccc ttaatctcca aagcaatgat gaataccgtg agacagacga tcattgtcat
     1441 tactttgtat attgccacca gtttcccatt tatcggaagt atgctctggg caacgtggga
     1501 tcctaaagct ttcactttgc cattttttac tggggctgct ttcaccattc taagtctgct
     1561 aaatagtctc acaagttgcg tcaacccttg gatttacttt gcattcaaca aggaattacg
     1621 tggagcgttg acaaattttt tctacagaaa gaaagactcg ttgaattatg atatagatgc
     1681 acgtcaaaat gtgtcagatg tagcctctac gacgtcgtcg tttatttcta gaatttcacg
     1741 acttgcgagt tcaaagatat tcggcacggt accgcgggcc cgggatccac cggtcgccac
     1801 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1861 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1921 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1981 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2041 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2101 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2161 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2221 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2281 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2341 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2401 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2461 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2521 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2581 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2641 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2701 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2761 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2821 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2881 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2941 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3001 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3061 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3121 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3181 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3241 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3301 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3361 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3421 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3481 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3541 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3601 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3661 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3721 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3781 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3841 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3901 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3961 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4021 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4081 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4141 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4201 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4261 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4321 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4381 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4441 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4501 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4561 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4621 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4681 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4741 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4801 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4861 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4921 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4981 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5821 cctgattctg tggataaccg tattaccgcc atgcat
//
