LOCUS       sequence-211554-        6583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A zebrafish insulin promoter containing vector with a second 
            multiple cloning sites. Contains red eye marker, and is flanked with
            I-SceI sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  AUTHORS   Singh SP, Janjuha S, Hartmann T, Kayisoglu O, Konantz J, Birke S, 
            Murawala P, Alfar EA, Murata K, Eugster A, Tsuji N, Morrissey ER, 
            Brand M, Ninov N
  TITLE     Different developmental histories of beta-cells generate functional 
            and proliferative heterogeneity during islet growth.
  JOURNAL   Nat Commun. 2017 Sep 22;8(1):664. doi: 10.1038/s41467-017-00461-3.
  PUBMED    28939870
REFERENCE   2  (bases 1 to 6583)
  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: "2017-09-22"; volume: "8(1):664. doi"; pages: " 
            10.1038/s41467-017-00461-3"
COMMENT     Sequence Label: ins:MCS2; cryaa:RFP
FEATURES             Location/Qualifiers
     source          1..6583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    37..61
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     intron          1256..1828
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_signal    2033..2154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2070..2089)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2124..2143
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2197..2404
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2393..2410
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2428..3138)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2706..2725)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2975..2993
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3722..3742)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3722..3740)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3773..3790)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3773..3789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3782..3804)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3930..4385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4279..4298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4412..4516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4517..4585,4586..5377)
                     /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     complement(4735..4754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5548..6136
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6037..6056
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6290..6307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6424..6445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6460..6465,6466..6483,6484..6490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6498..6514
                     /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)."
     primer_bind     6503..6525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6522..6538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6522..6538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctcatcacaa gtttgtacaa aaaagcaggc
       61 tccgcggccg cccccttcac cacgcgtaag ctttgtttgt accataataa gctcattata
      121 taatatttaa aatgatactg catcattaat tgatatgttt tctagttatt tttaaatatt
      181 agcaactaaa ttaggaaatt atccatagta aatttaatgt aatattattt gttatattta
      241 acttcagccc acagtctagt ttagttgttg gtcccttatg agaaaaaatt tgggcacctc
      301 tgctctagtg cttgtaaaca tttttgagta tctatagttg aacatgaaag catttttttt
      361 tttttttttt taagaaaatg aagaaagcta aaaacctgta tccactgact tccattaaaa
      421 aaagcaaata ctagggatgt ctatggtgtc ttcttttgtg ttcaacaaat aagcagaaag
      481 taactgctga cagaacatta cattttgtgt gaatcccctt atattgtgtt aaaaactgta
      541 gagaaatcat ctaataataa gaaattatca agaaataaac agaaaatgat ctaatagtat
      601 actataaaat ctttcaaaat ccaggccact accatatgac taaaaccaca catattatgc
      661 tttaaaaaaa gtcacaatga agacgtcaca aacgttcctg aagactaatg actcgtctaa
      721 ccccatctta ccttctgctg tttcactcaa agaaaagcca ccagctgtct ctgtgatctg
      781 ctcattctgt gttaacttgt gttctcaaca attaacaatt aaccaaaagc ctctggagct
      841 gcgtgagcat cacgatgtgt ttgtcttctg tgagcgtggc agtgtccacg aagtcaatgt
      901 gaattggttg gccaaacaca gctaatgtgg catgtgtgat tggacagcca cttaaaagtg
      961 ttaaagtgtg tttacgcaca ggattggttt tccgctgaat caggcaaaac cagcagaacc
     1021 gatctaaact cgagtataaa agtgagtagg agatgctatt cgtcccaaaa catctccacc
     1081 accatctcca ccattcctcg cctctgcttc gagaacaggt gagtgtcgag cgggatggta
     1141 aatctacaga gaatgcggag tgtagcttgt gtacatgttt ttgattaaca gagattgtat
     1201 gtgtgtgttt gtgtcagtgt gaaagggtgg gcgcgccgat cctgagaact tccgggtgag
     1261 tttggggacc cttgattgtt ctttcttttt cgctattgta aaattcatgt tatatggagg
     1321 gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt cccttgtatc accatggacc
     1381 ctcatgataa ttttgtttct ttcactttct actctgttga caaccattgt ctcctcttat
     1441 tttcttttca ttttctgtaa ctttttcgtt aaactttagc ttgcatttgt aacgaatttt
     1501 taaattcact tttgtttatt tgtcagattg taagtacttt ctctaatcac ttttttttca
     1561 aggcaatcag ggtatattat attgtacttc agcacagttt tagagaacaa ttgttataat
     1621 taaatgataa ggtagaatat ttctgcatat aaattctggc tggcgtggaa atattcttat
     1681 tggtagaaac aactacaccc tggtcatcat cctgcctttc tctttatggt tacaatgata
     1741 tacactgttt gagatgagga taaaatactc tgagtccaaa ccgggcccct ctgctaacca
     1801 tgttcatgcc ttcttctctt tcctacagct cctgggcaac gtgctggttg ttgtgctgtc
     1861 tcatcatttt ggcaaagaat tactagtgga tccgatatcg aattcttaat taaggccgcg
     1921 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1981 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaatcgag
     2161 ggggggcccg gtacccaccg cggtggcggc gatctctccc cagcatgcct gctattgtct
     2221 tcccaatcct cccccttgct gtcctgcccc aacccacccc ccagaataga atgacaccta
     2281 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
     2341 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
     2401 acagtcgagg ctgatcagcg ggccgcttta cttgtacagc tcgtccatgc cgccggtgga
     2461 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     2521 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     2581 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     2641 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     2701 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     2761 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     2821 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     2881 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     2941 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     3001 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     3061 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     3121 ctcgcccttg ctcaccatgt tgtcagacct ggtaactcct tactgtaact tattgcacct
     3181 gatgaacaaa agatgtgaac tgaacgcaag taggaggcta atagaggcta gtacagtaca
     3241 aagatcccca aatgatgtgt gccggcctta gtggaggcag acccctggtt atatacctgt
     3301 gctgcagaaa gaggggctta caaatggatc tctctggaat ggcattatct catggtggga
     3361 acagtgttgt cagcagaatc agcggggtgg actctgacct gagaatgaca gtctagcact
     3421 gaatggctca gacaatactc ttgtggccaa ctatcaccgg aaaataagtg cctgccaaaa
     3481 caaaccagac cagatgaagc tacatggctg tagaacgtca gtatttaatg tgttgtggcc
     3541 acttggttca ccagatttaa gagctaattt ttttaaatat cacagataat caatcgcttt
     3601 ttttttttct tgactgaaaa gcttagactg gaaaattctt tgaaaaagtt tgcactacag
     3661 aaaataaata cataacatcc tgcactgaat gcacactatt aatggtaccc agcttttgtt
     3721 ccctttagtg agggttaatt gcgcgctagg gataacaggg taatacgcgc tcactggccg
     3781 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3841 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3901 aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt
     3961 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4081 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4141 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4201 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4261 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4321 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4381 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4501 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4561 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4621 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4681 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4741 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4801 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4861 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4921 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4981 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5041 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5101 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5161 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5221 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5281 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5341 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5401 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5461 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5521 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5581 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5641 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5701 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5761 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5821 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5881 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5941 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6001 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6061 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6541 tgattacgcc aagcgcgtta gggataacag ggtaatgcgc gcg
//
