LOCUS       sequence-211549-        6584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A zebrafish insulin promoter containing vector with 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 6584)
  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 6584)
  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:MCS; cryaa:RFP
FEATURES             Location/Qualifiers
     source          1..6584
                     /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    2034..2155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2125..2144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    2198..2405
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2394..2411
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2429..3139)
                     /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(2707..2726)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2976..2994
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3723..3743)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3723..3741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3774..3791)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3774..3790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3783..3805)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3931..4386)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4089)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4280..4299
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4413..4517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4518..4586,4587..5378)
                     /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(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6291..6308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6425..6446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6461..6466,6467..6484,6485..6491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6499..6515
                     /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     6504..6526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6523..6539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6523..6539
                     /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 tcgatatcgc tagcatgcat gtcgacttaa ttaaggccgc
     1921 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     1981 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2041 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaatcga
     2161 gggggggccc ggtacccacc gcggtggcgg cgatctctcc ccagcatgcc tgctattgtc
     2221 ttcccaatcc tcccccttgc tgtcctgccc caacccaccc cccagaatag aatgacacct
     2281 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     2341 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     2401 cacagtcgag gctgatcagc gggccgcttt acttgtacag ctcgtccatg ccgccggtgg
     2461 agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     2521 tgatgtccaa cttgatgttg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct
     2581 tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct
     2641 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc
     2701 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     2761 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     2821 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcaagt
     2881 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg
     2941 acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     3001 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     3061 cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct
     3121 cctcgccctt gctcaccatg ttgtcagacc tggtaactcc ttactgtaac ttattgcacc
     3181 tgatgaacaa aagatgtgaa ctgaacgcaa gtaggaggct aatagaggct agtacagtac
     3241 aaagatcccc aaatgatgtg tgccggcctt agtggaggca gacccctggt tatatacctg
     3301 tgctgcagaa agaggggctt acaaatggat ctctctggaa tggcattatc tcatggtggg
     3361 aacagtgttg tcagcagaat cagcggggtg gactctgacc tgagaatgac agtctagcac
     3421 tgaatggctc agacaatact cttgtggcca actatcaccg gaaaataagt gcctgccaaa
     3481 acaaaccaga ccagatgaag ctacatggct gtagaacgtc agtatttaat gtgttgtggc
     3541 cacttggttc accagattta agagctaatt tttttaaata tcacagataa tcaatcgctt
     3601 tttttttttc ttgactgaaa agcttagact ggaaaattct ttgaaaaagt ttgcactaca
     3661 gaaaataaat acataacatc ctgcactgaa tgcacactat taatggtacc cagcttttgt
     3721 tccctttagt gagggttaat tgcgcgctag ggataacagg gtaatacgcg ctcactggcc
     3781 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3841 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3901 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     3961 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4021 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4081 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4141 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4201 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4261 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4321 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4381 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6241 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6301 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6361 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6421 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6481 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6541 atgattacgc caagcgcgtt agggataaca gggtaatgcg cgcg
//
